Malignant Melanoma Healed with Natural Therapies

How To Prevent Skin Cancer

How To Prevent Skin Cancer

Complete Guide to Preventing Skin Cancer. We all know enough to fear the name, just as we do the words tumor and malignant. But apart from that, most of us know very little at all about cancer, especially skin cancer in itself. If I were to ask you to tell me about skin cancer right now, what would you say? Apart from the fact that its a cancer on the skin, that is.

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How I Survived Malignant Melanom

By The Time You've Finished Reading How I Survived Melanoma Skin Cancer Seven Survivors Tell Their Stories. You'll Feel Like A New Person, with A New, More Positive Outlook! You will learn: 1. How do I know if I have melanoma? What are the signs and symptoms? I wanted to know why the doctor was so concerned when she looked at that little mole on my forearm. What was it that looked so sinister? How worried should I be? Was the doctor over-reacting? 2. What tests will the doctor carry out to see if I have melanoma? Will they be able to tell me on the spot if there is a problem? Or will I have to wait for days, fretting about whats going on? 3. How curable is melanoma? If they do tell me its melanoma, what exactly does that mean? Is it a death sentence? Will they tell me You have 12 months to live. Get your life in order and prepare for the worst.? 4. What are the stages of the disease? The reading Id done said that there were different stages of melanoma. What are the symptoms of each stage? What are the survival rates of each stage? If I had a later stage melanoma, wouldnt I know about it? Wouldnt I actually feel like I was sick? 5. How quickly does the disease progress or spread? Should I have gone to the doctor sooner? Id noticed the mole changing over about 3 months. Was this delay critical? 6. How is melanoma normally treated? Would I have to go through chemotherapy and radiation treatment? If so, for how long? What are the odds of curing the disease using these treatments? How extensive is any surgery likely to be? How big will the scars be? 7. What are the common side effects of the treatments? Would I lose my hair? Would I become sterile? What else could I expect? 8. What alternative treatments are available? Id heard of people going on special macro-biotic diets. Id seen lots of herbal remedies on the internet. Which of these are proven and documented, and which ones are snake oil? Is it possible to combine alternative treatments with surgical other western treatments? How do I find a doctor that is open to using both alternative and western treatments? 9. What are the latest treatments being developed, and who is carrying out clinical trials of these new treatments?

How I Survived Malignant Melanom Summary

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Allogeneic melanoma lysates for the treatment of disseminated melanoma

We began our studies with allogeneic melanoma lysates late in 1985, in patients with disseminated melanoma. This was done primarily for ethical reasons, because we did not know what the toxicity of this form of treatment would be. That would preclude its use in patients with minimal residual disease the group that might best benefit from vac-cination until phase I data showed that toxicity was acceptable. We should note that the use of this admittedly crude whole-cell vaccine, together with an adjuvant that may further have obscured the effects of the tumor preparation itself, was a decision we made deliberately after some experience with extracts of autologous cells given without adjuvant our initial attempt to use a better defined, individual-specific antigen. The failure to generate either an immune response or a clinical response in a small group of patients led us to the diametrically opposite approach whole-cell lysates with antigens that gave the patients the choice of which to...

Other genetic factors and skin cancer

An increasing number of studies have recently reported on associations between various candidate genes and skin cancer. Most plausible are reports of associations between genes involved in DNA repair and skin cancer. Patients with xeroderma pigmentosum (OMIM 278730), an autosomal recessive disorder characterized by mutation in one of a number of genes involved in nucleotide excision repair, have a grossly elevated risk of both melanoma and NMSC (often presenting in early childhood with multiple tumors). Various studies have recently reported associations between some of the genes implicated in xeroderma pigmento-sum and skin cancer (Dybdahl et al., 1999 Tomescu et al., 2001 Winsey et al., 2000), whilst other studies have reported on associations between polymorphisms in pathways that may play a role in oxidative damage control in skin in response to UVR (such as the glutathione transferases) (Lear et al., 1997 Ramsay et al., 2001) or skin immunity (tumour necrosis factor a) (Hajeer et...

Uveal melanoma of choroid and ciliary body

Malignant melanoma of choroid. W Figure 37.1. Malignant melanoma of choroid. W Figure 37.2. Malignant melanoma of choroid. W Figure 37.2. Malignant melanoma of choroid. W Figure 37.3. Malignant melanoma of choroid. W Figure 37.3. Malignant melanoma of choroid. W Figure 37.4. Malignant melanoma of choroid. W Figure 37.4. Malignant melanoma of choroid. W

The MC1R and human pigmentation and skin cancer

In 1995 we showed that sequence variation at the MC1R was common in man and that particular variants were associated with red hair and ''pale'' skin (Valverde et al., 1995). Such individuals are characterized clinically by a tendency to burn rather than tan in response to UVR, and pale skin on sites protected from the sun (such as the buttock). This clustering of phenotypic characteristics is known to be associated with an elevated risk of most forms of skin cancer (Rees, 2002c). Red hair approximates to an autosomal recessive trait although the penetrance depends on which MC1R variants are present (typically penetrance is around 0.85). This bold statement ignores some of the subtleties of phenotype hair color changes with age from childhood through early and late adulthood, and different body sites frequently have different colored hair. Given that the MC1R is a determinant of pigmentary phenotype and that pigmentary phenotype is a risk factor for both melanoma and NMSC it is not...

Mucosal Melanomasspecial Considerations

Mucosal melanomas of the head and neck are rare, with only about 1000 cases reported. Of more than 84,000 melanoma cases in the National Cancer Data Base, only 1.3 were mucosal, and 55 of that subgroup arose in the head and neck (15). The majority of head and neck mucosal melanomas are sinonasal in origin, and the others are primarily from the oral mucosa. Among the sinonasal tumors, 81 arise in the nose and 19 in the sinuses, although the size of some of these tumors makes it difficult to be sure of the site of origin. Most of the oral lesions arise from the hard palate and maxillary alveolar ridge. Melanomas arising in other head and neck mucosal sites such as the larynx, pharynx, and cervical esophagus have been reported, but are quite rare. A melanoma arising in the oropharynx is shown in Figure 3. The oral lesions are often detected by healthcare personnel as a dark spot in the mouth, but the sinonasal lesions are often only apparent after symptoms such as epistaxis or nasal...

Is There an Association Between Levodopa Therapy and Melanoma

The connection between levodopa therapy, PD, and malignant melanoma has been a matter of debate for three decades. It originally derived from the biochemistry of the drug. Levodopa is a substrate for the development of dopamine, which, in turn, develops into neuromelanin in CNS nigral neurons. It is the dopaquinones derived from levodopa that are oxidized to form neuromelanin in these cells (126). Hence, it has been proposed that levodopa may also affect the activity of melanocytes in the skin, possibly promoting malignant transformation, although this connection has never been proven. In addition, it is now known that 70 of melanoma cases in the general population appear to be connected with a genetic mutation unrelated to PD. Thus, it would seem unlikely that there would be a connection between PD, lev-odopa, and melanoma. Nevertheless, reports of melanoma in patients treated in the 1970s led the FDA to require language in the package insert that cautions against the use of levodopa...

Glioma and Melanoma

Yamanaka and colleagues have used SFV replicons for multiantigen immunotherapy in the 203 glioma and B16 melanoma models derived from C57BL 6 mice (54). Tumor cDNA was generated from tumor mRNA and cloned into the SFV replicon constructs yielding SFVcDNA-RNA (54). This mixture of SFV replicons expressing a wide array of tumor-derived heterologous proteins was incubated with syngeneic, Day 6, bone marrow-derived DCs that were then used as the immunogen via three intraperitoneal injections at 1-wk intervals. Splenocytes from immunized animals were evaluated for CTL responses and the SFV replicon-loaded DCs yielded substantial tumor-specific CTL responses, approximately twofold greater specific cytolysis at each effector to target ratio compared to splenocytes from animals immunized with tumor RNA or tumor lysate-pulsed DCs. In tumor protection models, tumor outgrowth was delayed with the SFV-cDNA replicon-loaded DC immunizations and 50 full protection was demonstrated using intracranial...

Skin Cancer

Balch CM, Buzaid AC, Soong SJ, Atkins MB, Cascinelli N, Colt DG, Fleming ID, Gershenwald JE, Houghton A Jr, Kirkwood JK, McMasters KM, Mihn MF, Morton DL, Reintgen DS, Ross MI, Sober A, Thompson JA, Thompson JF. Final version of the American Joint Committee on cancer staging system for cutaneous melanoma. J Clin Oncol 2001 19 (16) 3635-3648. Banerjee SS, Harris M. Morphological and immunophenotypic variations in malignant melanoma. Histopathology 2000 36 387-402. Breslow A. Thickness, cross-sectional areas and depth of invasion in the prognosis of cutaneous melanoma. Ann Surg 1970 172 902-908. Clark WH Jr. A classification of malignant melanoma in man correlated with histogenesis and biological behaviour. In Montagna W, Hu F (eds).Advances in Biology of the Skin. Vol. 8. Pergamon Press. London 1967 621-627. Cochran AJ, Bailly C, Cook M, Crotty K, Mihm M, Mooi W, Sagebiel R. Recommendations for the reporting of tissues removed as part of the surgical treatment of cutaneous melanoma....

Non Skin Cancers

Significantly increased relative risk not only for non-melanoma skin cancer but also for buccal cancer and primary liver cancer. Whether this increased relative risk was correlated with alcoholism and or smoking could not be ascertained in this study. The data suggest a co-occurrence of HS and cancer but these observations await confirmation in other HS populations. If confirmed it may give direction to further aetio-logical and pathogenic research in HS.

Skin cancers

There are two main types of skin cancer melanoma, arising from the neural crest derived melanocytes and basal cell carcinoma and squa-mous cell carcinoma derived from keratinocytes (the latter two referred to as non-melanoma skin cancer, NMSC) (Rees, 2002b). This broad distinction into two types is useful NMSC is extremely common and case-fatality extremely low (

Melanoma

Rates of melanoma vary widely geographically, with the major determinant of risk being skin color. The relative incidence of subtypes of melanoma differs across racial groups with one rare subtype termed ''acral lentiginous melanoma'' occurring in all racial groups (Hudson and Krige, 1993 Stevens et al., 1990 Jimbow and Kukita, 1984) but the commoner subtypes (superficial spreading melanoma, nodular melanoma and lentigo maligna) occur essentially only in white persons. For these latter subtypes and among white populations, the highest incidence is in countries nearest to the equator, leading to the hypothesis that sun exposure is the major environmental determinant of melanoma risk (Armstrong, 1988). Australia and New Zealand have the highest rates worldwide (Jones et al., 1999 Parkin et al., 1997). Variation in skin color explains much of the variation within and between geographical locations. For instance, in Hawaii, the melanoma incidence rates for people of European descent are...

Michael A Morse MD Timothy M Clay PhD and H Kim Lyerly MD

Interest in activating the immune system to control tumors dates to more than 100 yr ago and is often attributed to William B. Coley who used the inflammatory response to bacterial products as a form of immunotherapy (1). This initiated the era of attempts to nonspecifically activate the immune system with the hope that tumors would be targeted as part of the overall, predominantly inflammatory, response. Following the abandonment of the crude bacterial products in Coley's toxin, other bacteria or their cell wall products such as bacille Calmette-Guein (BCG), Corynebacterium parvum, nocardia rubra, OK-432, and others have been tested. In some circumstances, these inflammatory activators have had success as antitumor agents. For example, BCG has activity in superficial bladder cancer in situ (2). Later, cytokines such as interleukin (IL)-2 that could activate T and natural killer (NK) cells were extensively tested and demonstrated activity in renal cell carcinoma and melanoma. In the...

Ancillary Techniques in Histopathology Specimens

Immunohistochemistry The surgical pathologist's second H and E is invaluable in assessing tumour type, prognosis and treatment, e.g., carcinoma (cytokeratins) versus malignant lymphoma (CD45) and malignant melanoma (S100), or better prognostic and hormone responsive breast cancer (oestrogen receptor positive). Tumour antigenic profile is also of use in specifying the site of origin for a metastasis, e.g., prostate carcinoma (PSA positive). Electron microscopy Valuable in renal biopsy diagnosis, and tumours where morphology and immunohistochemistry are inconclusive, e.g., malignant melanoma (pre- melanosomes) and neuroendocrine carcinoma (neurosecretory granules). Quantitative methods Prognostic indicators include the Breslow depth of invasion in malignant melanoma, muscle fibre typing and diameter in myopathies and the mitotic activity index in breast carcinoma.

Molecular Correlates Of Germ Cell Tumor Development And Function In Humans

C-Kit is a transmembrane glycoprotein (molecular weight, 145Kd) produced by the c-kit gene and is a member of the receptor tyrosine kinases. It is related to the receptors for platelet-derived growth factor, macrophage colony-stimulating factor, and the flt 3 ligand.70 In mice, its natural ligand is the Steel factor or the stem cell factor, and murine studies suggest that the function of Kit tyrosine kinase is important in the development of the interstitial cells of Cajal, mast cells, and normal hematopoiesis71 and in the development and maturation of spermatogonia.43 Curiously, c-Kit expression or mutation has been observed in a range of human malignancies, including mast cell leukemia, gastrointestinal stromal tumors, melanoma, and seminoma.7,72,73 Tian and colleagues7 found activating mutations of c-kit in about 10 of seminomas but in no instances of NSGCT, but it should be noted that these workers studied only exons 11 and 17. The commonality of c-kit mutations in melanoma, GCTs,...

Antiepileptic Drugs In Neurooncology Patients

Approximately 25-30 per cent of the adults and 10-15 per cent of the children with primary or metastatic brain tumors will experience a seizure during the course of their illness 1-4 . Seizure incidence is even higher in patients with primary glioma, particularly low-grade glioma and in patients with metastases from melanoma 3,5-10 . Although routine prophylaxis with AEDs is not recommended for patients with a brain tumor who have not had a seizure, patients in the community are often on an AED, usually phenytoin 2 . This practice can lead to

Chemotherapy And Boron Neutron Capture In Rat Brain Tumor Models

Neutron flux at the Brookhaven National Laboratory Medical reactor appeared curative for rats bearing 9L gliosarcoma. On the other hand, the F98 tumor was refractory to therapy. Barth et al., 49 went on to determine whether the efficacy of boron neutron capture therapy could be increased by combination with standard radiation therapy. F98 glioma-bearing rats treated with intracarotid or intravenous sodium borocaptate (30 mg kg) plus p-boronophenylalanine (250 mg kg followed 2.5 h later by exposure to a neutron flux and 7-10 days later by standard radiation therapy to a total dose of 15 Gray had survival times (61-53 days) similar to those of rats that did not receive the standard radiation therapy (52-40 days). However, animals in the same study bearing MRA 27 melanoma showed a significantly enhanced survival when boron neutron capture therapy was followed with conventional radiation therapy.

Identification of Class I Peptides

Once a class I specific peptide has been identified, one must determine whether that peptide can be used to generate T cells capable of responding to and lysing HLA-matched tumor that expresses the antigen of interest. These studies may be performed by using peptide to sensitize T cells in vitro. However, in vitro propagation of human lymphocytes is labor intensive and operator dependent. In addition, establishing T-cell lines using lymphocytes derived from patients with cancer may be made more difficult due to potential inherent defects in the T-cell response in tumor-bearing hosts. For these reasons, another approach has been developed to rapidly evaluate the utility of predicted class I epitopes, active immunization of HLA-A2 transgenic mice. Vaccinating mice with class I peptides allows a more robust immune response to be generated to a particular tumor antigen than the response found endogenously in cancer-bearing patients. Moreover, initiating in vitro culture with a robust...

From The Laboratory To The Clinic Oncolytic Herpes Simplex Viruses Appear Safe In Trials But Just How Effective Will

Adverse events, such as acute toxicity, viral shedding, clinically evident reactivation or encephalitis attributable to virus inoculation or replication were observed in either study at any of the doses, which escalated from 103-105 pfu with 1716 and 1 x 1063 x 109 with G207 (16,17). Patient deaths were primarily the reuslt of progressive cancer, with one report of radiation necrosis. Two of the G207 treated patients were alive as of December, 2003 (77). In addition, both studies were unable to establish a maximum tolerated dose (16,17). More recently, evidence has been obtained that is reportedly consistent with replication of 1716 within tumor samples over time, supporting the concept of tumor destruction by viral oncolysis (78). Finally, multiple injections of 1716 directly into skin lesions of a limited number of melanoma patients have proceeded without any harmful effects (79). A phase Ib II study involving G207 is currently underway open to patients with recurrent glioblastoma...

Gangliosides GM2 GD2 GD3 and Fucosyl GM1

Gangliosides are sialic acid-containing glycolipids that are expressed at the cell surface with their lipid (ceramide) moiety incorporated into the cell-surface lipid bilayer. Most gangliosides considered as potential targets for cancer therapy are expressed primarily in tissues and tumors of neuroectodermal origin. This is true for the melanoma, sarcoma, neuroblastoma, and SCLC antigens GM2, GD2, and GD3, and the SCLC antigen fucosyl GM1. The structures of these antigens are shown in Fig. 2. Surprisingly, however,

Tumor Targeting Of Alphavirus Vectors

Yet another approach to obtain tumor selective transfection was attained by encapsulation of recombinant SFV particles in liposomes (40). By this procedure, targeted gene delivery to human LnCaP prostate tumors implanted in severe combined immunodeficiency (SCID) mice was achieved after systemic delivery of encapsulated SFV-LacZ particles. Systemic administration of encapsulated SFV particles expressing the p40 and p35 subunits of IL-12 to SCID mice with human Panc-1 pancreatic tumors resulted in statistically significant reduction in tumor growth after a single injection (41). Furthermore, an initial phase I study on advanced melanoma and kidney carcinoma patients demonstrated the safe use of this SFV vector in humans. In this study the maximum tolerated dose (MTD) for encapsulated SFV-IL-12 particles was 3 x 109 particles m2, which might seem relatively low compared with doses for other viruses, usually administered in the range of 1011 particles. However, in this case the dose is...

Cytokine expression driven by pox vectors

Both orthopox (vaccinia) and avipox (fowlpox and canarypox) vectors have been used for the expression of GM-CSF. Previous studies have shown that direct injection of rV-GM-CSF into tumor lesions will enhance antitumor effects both in experimental animals (73) and in clinical studies in melanoma patients (74). On the other hand, fowlpox expresses transgenes for 14-21 d. A recent study in mice has compared the biological activity of recombinant GM-CSF with that of fowlpox expressing GM-CSF (40). A single injection of rF-GM-CSF led to significantly higher numbers of APCs, particularly DCs, in regional lymph nodes when compared with four daily injections of GM-CSF. In

Proteasome Inhibitors

The first in vivo demonstration of the antitumor activity of proteasome inhibitors involved treatment of Burkitt's lymphoma xenografts with PS-341, since renamed Bortezomib 173 . A screen of tumor cell lines isolated from 9 different cancer types (brain, colon, melanoma, prostate, lung, breast, renal, and ovarian) showed that Bortezomib had broad antitumor activity both in vitro and in vivo 174,175 . Additional studies found that transformed cells were selectively sensitive to the cytotoxic effects of protea-some inhibition when compared to normal cells in culture 173,176-179 . In vivo effects were detected in prostate tumor xenograft and murine mammary carcinoma models 180 . trials evaluated the use of bortezomib in the treatment of both chemoresistant and recurring solid tumors (hepatocellular carcinoma, melanoma, metastatic breast cancer, metastatic colorectal carcinoma, meta-static renal cell carcinoma, neuroendocrine tumors, ovarian carcinoma, non-small cell lung carcinoma, and...

Melanocortin 1 Receptor

The melanocortin 1 receptor (MC1R) is a major determinant of hair and skin pigmentation. Extensive case-control studies and studies on familial melanoma showed that genetic variations in MC1R play crucial roles in the development of both familial and sporadic melanoma.5152 More importantly, certain variants of the MC1R were associated with development of melanoma, independent of pigment synthesis. Therefore, apart from pigment synthesis, the MC1R is involved in hitherto unknown tumorigenic signal transduction pathways. Whether the genetic variants of MC1R harbor constitutively active properties, thus providing a causative link to aberrant proliferation, is yet to be determined.

Global Analysis of Gene Expression

In a few cases, it has been possible to confirm functional hypotheses first suggested by expression analysis. For example, the gene encoding RhoC (ARHC) was identified as an expression correlate of tumor metastasis in a melanoma model critically, blockade of RhoC diminished metastasis and activation enhanced metastasis in this model.57 As yet, few such functional hypotheses have been validated, due to the relative mismatch between our ability to rapidly generate such hypotheses using expression microarrays and our more limited ability to test them in the laboratory. However, methods for systematically testing gene function on a global scale are rapidly evolving and are discussed briefly in the next section.

Neoplastic Conditions

Others Low-grade sinonasal adenocarcinoma, olfactory neuroblastoma, malignant melanoma, small cell neuroendocrine carcinoma, sinonasal undifferentiated carcinoma, rhabdomyosarcoma, chondrosarcoma and chordoma are all uncommon. Prognosis outcome depends on the histological type of tumour as well as the extent of spread. Most lesions are advanced at presentation although lymph node metastasis with carcinomas is relatively infrequent. Local recurrence is a common problem in spite of radical surgery and radiotherapy. Melanomas, small cell neuroendocrine carcinomas and sinonasal undifferentiated carcinomas are particularly aggressive but five-year survival is the norm with adenoid cystic carcinomas. In certain subtypes, such as intestinal-type sinonasal adenocarcinoma and olfactory neuroblastoma, grading based on the degree of differentiation is important in that low-grade lesions do well while high-grade lesions do badly. Around 20 five-year survival is customary.

Summary And Conclusions

Our studies and those of others (11, 25) indicated that IL-2R and endogenous IL-2 are ubiquitously expressed in human carcinomas both in culture and in situ. Data available from various laboratories provide evidence for the presence of IL-2R on melanoma cell lines and other tumour cell lines. Furthermore, the IL-2 IL-2R pathway appears to be operative in normal non-hematopoietic tissues, e.g., fibroblasts and keratinocytes (5, 26). Thus, this pathway appears to be active in vivo and in 6. Rimoldi D, Salvi S, Hartmann F, Schreyer M, Blum S, Zografos L, Plaissance S, Azzarone B, Carrel S. Expression of IL-2 receptors in human melanoma cells. Anticancer Res 1993 13 555-564 12. Alileche A, Plaisance S, Han DS, Rubinstein E, Mingari C, Bellomo R, Jasmin C, Azzarone B. Human melanoma cell line M14 secretes a functional interleukin-2. Oncogene 1993 8 1791-1796 29. Atkins MB. Interleukin-2 in metastatic melanoma establishing a role. Cancer J Sci Am 1997 3 57-58

Oncogenic Capacity Of The Jakstat Signaling Pathway

Extensive data describe activated Stat3 and Stat5 in tumors. Activating mutations of Stat3 and Stat5 and specific inhibition of Stat3 and Stat5 by gene deletion, antisense oligonucleotides, or dominant negative approaches have highlighted the importance of Stat3 and Stat5 in tumor formation. Mutations of Stat3 that allow spontaneous dimer-ization of the monomers in the absence of interactions between phosphorylated tyrosines and SH2 domains are sufficient to cause transformation and induce tumor formation in nude mice (98). Inhibition of Stat3 signaling using antisense Stat3 or by a Jak-selective tyrosine kinase inhibitor, AG490, restored the sensitivity of cells from patients with large granular lymphocyte (LGL) leukemia to Fas-mediated apoptosis (99). Downregulation of Fas correlates with an increase in metastatic potential and resistance of tumors to chemically and physically induced apoptosis. This effect is mediated, at least in part, by an interaction between Stat3 and c-jun,...

The CD40CD40L Pathway

CD40 is a member of the tumor necrosis factor receptor (TNFR) family (62-65) broadly expressed on antigen-presenting cells (APCs), B cells, epithelial cells, and endothelial cells (66). Engagement of CD40 on APCs by its ligand CD40L (CD154) on responding T cells augments the resulting immune response (66). Exogenous activation of the CD40 pathway by agonist antibodies is capable of substituting for T-cell help (67), and augments both humoral and CTL responses (68). Manipulation of the CD40 pathway by engineering chronic lymphocytic leukemia (CLL) cells to overexpress CD40L by adenoviral gene transfer induces the expression of multiple costimulatory molecules. This augments the antigen-presenting capacity of both CD40L-modified CLL cells and unmodified bystander CLL cells in vitro, enabling both of them to prime CTL specific for autologous CLL (69). CD40L modification also augments the antigen-presenting capacity of human multiple myeloma cells in vitro (70,71). The ability of...

Cancers Other Than Breast and Ovarian Cancer

Carriers families rectal Stomach bladder Pancreas Liver duct Uterine Pertonium tube Cervix Prostate Lymphoma cell skin Leukemia Melanoma combined Malignant Melanoma An increased risk of cutaneous malignant melanoma associated with having an affected family member was quantified (OR 2.69) in a study by Holman and Armstrong (124). It has been estimated that 8 to 12 of cases are attributable to inherited factors (11). There is a known association of malignant melanoma with a mutation in CDKN2 on chromosome 9 that codes for p16, another important regulator of the cell division cycle (125). Several studies have demonstrated an association of BRCA mutations and malignant melanoma. In 3728 individuals in 173 breast-ovarian cancer families with BRCA2 mutations, the BCLC (36) estimated a statistically significant RR for malignant melanoma of 2.58. However, a study by van Asperen et al. (126) noted no significant excess risk of malignant melanoma associated with BRCA mutations in first-degree...

Extent Of Local Tumour Spread

Carcinomas with a pushing border and prominent lymphocytic reaction are regarded as having a better prognosis than those with a diffusely irregular infiltrating margin and sparse lymphocytic reaction, e.g. col-orectal carcinoma, head and neck carcinoma, malignant melanoma, medullary carcinoma of breast, advanced gastric carcinoma. Malignant melanoma. Direct linear measurement (mm) and anatomical level of invasion of the vertical component are strong prognostic indicators. The TNM classification is applied to carcinoma only in the majority of tissues. Other qualifying malignant tumours are malignant mesothe-lioma, malignant melanoma, gestational trophoblastic tumours, germ cell tumours and retinoblastoma.

Dose and Schedule of Administration

Five vaccine dosage schedules have been tested and are summarized in Table 2. The animal tumor models, particularly the paper of Fujiwara et al. (32), prompted us to presensitize patients with DNFB by topical application of a 1 solution in acetone-corn oil, and this was done in schedules A, B, and C. Subsequently, the presensitization was found to be unnecessary (but not deleterious) for the induction of maximum DTH to autologous melanoma cells and it was omitted for schedules D and E. Melanoma 1) Measurable metastases 97a

Development of Delayed Type Hypersensitivity Responses 861 Method for DTH Testing

Patients were tested for DTH by modification of standard methodology (45). Cryopreserved melanoma cell suspensions and PBLs were thawed, washed, and irradiated (2500 cGy). DNP modification of melanoma cells and PBLs was performed as described above. Melanoma cells (1 x 106) and PBLs (3 x 106), each either DNP modified or unmodified, were suspended in Hanks balanced salt solution without serum, phenol red, or antibiotics and injected intradermally into the ventral forearm. The mean diameter of induration was measured after 48 h. A positive response was defined as maximum diameter of induration 5 mm. Patients were also skin tested with intermediate strength purified protein derivative (PPD)(5 TU). DTH testing was performed before the treatment program was initiated and at various times post-treatment. Analyses were performed by determining the maximum DTH response exhibited by each patient to each of the test reagents. Pretreatment positive DTH responses to autologous melanoma cells,...

Studies to investigate genes and environment

The Melanoma Genetics Consortium (Bishop et al., 2002) examined the risk (penetrance) for melanoma among CDKN2A mutation carriers. Carriers of particular germline mutations in CDKN2A are known to be at increased risk of melanoma. In this analysis, based on families with multiple persons with melanoma and a germline CDKN2A mutation, the overall risk of melanoma was estimated to be 62 by age 75 years. However, there was statistical evidence of a difference with carriers in Australia having a risk higher than that of the USA which, in turn, was higher than that in Europe. In the general population, for comparable time periods, the risk of melanoma by age 75 was at its highest in Australia in which it approached 2 . On an age-specific basis, the population rates in Australia were approximately 7 times higher than in Europe, while the rates in USA were 5 times higher than those in Europe. With the limited precision of the estimates of the penetrance of CDKN2A in the published analysis, the...

Clinical Effects of DNPModified Autologous Vaccine

We have reported the results of a series of clinical trials of patients with surgically incurable metastatic melanoma with measurable metastases (48) there were 97 patients, of whom 83 were evaluable (Group 1 in Table 3). Among the 83 evaluable patients there were 11 responses 2 complete, 4 partial, and 5 mixed 2 patients were judged to have stable disease. Both complete responses and two of the four partial responses occurred in patients with lung metastases. Response durations were as follows partial responses 5, 6, 8, and 47+ mo complete responses 12, 29 mo. Two examples of antitumor responses are provided below Patient no. 20254, a 77-yr-old man, presented simultaneously with a regional lymph node metastasis in the neck and 2-cm diameter mass in the lung adjacent to the cardiac border that increased in size over 2 mo observation. Five months after beginning DNP-vaccine treatment, the same mass was thought to be slightly smaller. The mass continued to slowly regress and by the 2-yr...

Importance of Dosage Schedule

Our demonstration that the development of DTH to unmodified melanoma cells was critical to its clinical effectiveness has allowed us to study the dose response of the vaccine using DTH as the response. An analysis of 284 patients (Groups 2-4 in Table 3) who were treated following resection of regional or distant metastases showed no significant association between the magnitude of DTH and the number of live (trypan blue-excluding) melanoma cells administered per dose over a dosage range of 0.5-25.0 x 106 It has become apparent that the variation in intensity of DTH responses to autologous melanoma cells among patients receiving different treatment schedules was likely to be explained by the timing of what we have called an induction dose of vaccine (62). This is defined as an intradermal injection of autologous melanoma cells without BCG, which was intended as a baseline test of DTH reactivity. Surprisingly, retrospective analysis showed that patients who received the induction dose...

Environment and high penetrance genes

Genes associated with high penetrance of cancer have been identified for breast cancer susceptibility (genes BRCA1, BRCA2 and TP53) (Chapters 15,16), colorectal cancer (APC germline mutations and mismatch repair genes, predominantly hMSH2 and hMLH1) (Chapter 17) and melanoma (CDKN2A) (Eeles et al., 2004). The risks of malignancy at one or more anatomical site are 0.60 or higher by age 75 years for mutations in each of these genes. Population-based studies suggest that approximately 1 in 500 of the general population carries a germline mutation in BRCA1 or BRCA2 (although some populations, such as Ashkenazim, have notably higher carrier frequencies. Carrier frequencies for germline mutations in the other predisposing genes may be marginally lower than for BRCA1 and BRCA2. Persons carrying such mutations represent a challenge for clinical management and an important group in which to examine the effects of environmental exposures as any exposures which are modifiable would be targets...

Studies in Ovarian and Renal Cell Carcinoma

There are no practical impediments to trials of the DNP-modified, autologous vaccine in other cancers. However, given melanoma's reputation as being particularly immunogenic among human malignancies, some would speculate that the applicability of this immunological trick would be limited. We have conducted two phase I trials in patients with ovarian cancer. In the first trial, 9 evaluable patients (10 total) with newly diagnosed adenocarcinoma of the ovary underwent standard debulking surgery plus six cycles of chemotherapy (taxol + platinum) prior to receiving DNP-vaccine. In the second trial, 13 evaluable patients (20 total) with bulky, chemotherapy-refractory disease were treated. We chose dosage schedule D (see Table 2) because it appeared to induce optimal DTH responses in the melanoma studies. Positive DTH (5-mm induration) to autologous DNP-modified tumor cells was elicited in 19 of 22 patients (median 14-mm induration). More importantly DTH to unmodified tumor cells was...

Other Clinical and Preclinical In Situ Cytokine Gene Therapy Approaches

Been inserted into a vaccinia virus that coexpressed the MUC-1 gene, which was then delivered as an intramuscular injection to men with advanced prostate cancer (73). No toxicity was observed and one patient that received three doses had some evidence of systemic immune effects. IL-15 was shown to contribute to the development of NK and antitumor response to prostate cancer in a xenograft model with PC-3 tumors (74). IL-18 may synergize with IL-12 (75) and although it has not been used in gene therapy strategies for prostate cancer the recombinant protein has been used in combination with IL-12 gene therapy in a bladder cancer model (76). The IL-24 gene also known as a melanoma differentiation associated gene 7 (MDA-7) has been used in preclinical studies for prostate cancer gene therapy (77).

Non Cancer Health Care and Health Maintenance

The transition off of primary cancer treatment is also a second opportunity to consider whether genetic assessment might be necessary. During an initial consultation, when taking the family history, a potential genetic predisposition may be detected. However, the patient may not pursue referral to a genetic counselor at that time because they are so overwhelmed by the new diagnosis of cancer and dealing with the treatment they will have to embark upon. The completion of treatment is another opportunity to review this issue and consider making a referral. The genetics of breast, ovarian, and colorectal cancers are best understood, but increasingly associations with other cancers such as pancreatic cancer and melanoma are being

Cytokine Modified Dendritic Cells

DCs are extremely efficient APCs that are widely dispersed in tissues and peripheral blood. Because they can be manipulated ex vivo and are perhaps the most specialized APCs they have been considered the prime candidates for cell mediated cancer therapy. A number of clinical trials using DC have been performed for prostate cancer (109-116) as well as other cancers (see ref. 117 for a review of the first 1000 trials). Although melanoma is the most commonly treated cancer using DC immunotherapy, GU cancers, notably, renal and prostate, are also being evaluated (118). Most of these studies involve DC primed with specific polypeptides that will bind MHC Class I or II molecules or with tumor lysates. Tumor derived mRNA transfected into DCs is being used to circumvent the challenge of lack of identity of TAAs. In current clinical trials DCs are most often injected intradermally or subcutaneously and less commonly intravenously or into a lymph node (118). A challenge with current clinical...

The Role Of Il11 In Tumour Metastasis

Human melanoma (A375M) and human breast cancer (MDA-MB-231) cells formed osteolytic bone metastasis in vivo when these tumour cells were injected into the left ventricles of BALB c nude mice (Figure 2) (17). These tumour cells promoted bone resorption in the in vitro neonatal murine calvaria organ culture system by indirectly stimulating the production of a bone resorption-inducing factor (or factors) from human osteoblast-like Saos-2 cells. In the course of examining the factors, we found that Saos-2 cells, in response to the interaction with tumour cells, produced a factor that promoted the proliferation of T10 cells. Because the T10 cell growth was IL-6 and IL-11 dependent (24), we checked the concentrations of both in the Saos-2-conditioned medium. We found that this conditioned medium stimulated the production of IL-11, but not IL-6, from Saos-2 cells. Furthermore, we found that a specific anti-IL-11, but not anti-IL-6, antibody could neutralize the abilities to induce T10...

In Vivo Evidence for DCs As Strong Adjuvants

There is a large body of literature involving animal models of tumor immunity in which DCs loaded with tumor-associated antigens (TAAs) are able to induce protective antitumor responses. When tested, DCs can be superior to other vaccination strategies (24). There also are reports in which DC immunization produces significant therapeutic immunity to established tumors (reviewed in ref. 25). A number of trials have now utilized TAA-loaded DCs as vaccines in humans. Initial pioneering studies involved patients with lymphoma and melanoma (26,27). Some clinical and immune responses (T-cell proliferation and delayed-type hypersensitivity DTH ) without any major toxicity have been observed. More recent DC vaccination studies put further emphasis on the elicited immune responses and have included control antigens for CD4+ and CD8+ T-cell responses (28-31). Several of these studies are reviewed elsewhere in this handbook. Inclusion of control antigens helps to verify that the DCs are...

Assay Techniques 41 Identification of VSMCs

It may also be appropriate to confirm that cultures are von Willebrand factor negative if there is concern about endothelial cell contamination (for example, when isolating VSMCs from small blood vessels). Separate markers have also been described for microvascular SMCs (pericytes) such as 3G5 43 , but there is debate about the specificity of this marker 44 . The high molecular weight melanoma-associated antigen is also a marker for pericytes in vivo 28 .

Alkylating Agent Resistance

The use of MGMT point mutants to differentially protect the hematopoietic compartment while sensitizing tumors has also been reported in animal xenograft models (126-128). Clinical trials using gene transfer of MGMT have been proposed by several investigators, and one phase I trial in patients with advanced malignancies such as melanoma, sarcoma and other solid tumors is in progress (129). The objective of this trial is to protect bone marrow stem cells from the toxic effects of chemotherapy and select for MGMT-G156A transduced cells during treatment. This strategy is expected to result in less toxicity to bone marrow and blood cells while enriching for the number

Table 231 Different characteristics of benign and malignant tumours

Malignant disease accounts for 1 in 8 deaths of people under 35 years in Australia and 1 in every 4 (25 ) of deaths in those over 45 years. 1 Cancer is the only major cause of death in Australia that is increasing in both sexes. At current rates about 1 in 3 males and 1 in 4 females will develop a cancer, excluding non-melanoma skin cancers, by the age of 75. 1 Neoplasia, especially malignancy of the silent areas, can present as undifferentiated illness and be a real masquerade. The so-called 'silent' malignancies that pose a special problem include carcinoma of the ovary, kidney, caecum and ascending colon, liver (hepatoma), melanoma and haematological tissue.

Cytokines for Cancer Therapy

Cytokines such as interferons and interleukins are administered for cancer because of their broad-based immunostimulatory effects including generation of tumor-reactive lymphocytes (11). Interleukin-2 (IL-2), or aldesleukin, which is indicated for the treatment of adults with metastatic renal cell carcinoma and melanoma, is the most studied cytokine. IL-2 dose-dependently activates cellular immunity and causes release of other immune-boosting cytokines in vivo (11). Systemic cytokine therapy is generally limited by rapid degradation and elimination of the cytokine, the inability to achieve optimal concentrations in the tumor microenvironment, and dose-dependent toxicity, including life-threatening side effects such as vascular leak syndrome and orthostatic hypotension (12-15). Cytokine gene therapy, in which a cytokine gene (such as that for IL-2) is introduced into tumor cells, is being explored to overcome some of these limitations of systemic cytokine administration. However,...

AntiGD2IL2 Immunocytokine

The anti-GD2 immunocytokine hu14.18-IL-2 (EMD 273063) is currently in phase I clinical trials (18). This immunocytokine consists of recombinant human IL-2 and a humanized monoclonal antibody directed against human GD2, which is a disialogang-lioside antigen expressed by tumors of neuroectodermal origin including malignant melanoma, neuroblastoma, some sarcomas, and some small-cell lung carcinomas. Each immunocytokine molecule is composed of two cytokine molecules fused to each of the two heavy chains of a monoclonal antibody via a peptide linkage at the carboxyl terminus (Fig. 2). Preclinical studies with the anti-GD2-IL-2 fusion protein were conducted with a chimeric form (i.e., ch14.18-IL-2) as well as the humanized form that is EMD 273063 (i.e., hu14.18-IL-2). Early studies focused on establishing the biological functionality of the immunocytokine and on determining whether the fused protein retained full antibody and cytokine functions. That ch14.18-IL-2 retains its functionality...

Increasing Gene Expression

Replicons have proven to be powerful enhancements to DNA vaccination, and are capable of eliciting antibody and tumor protective responses at up to 1000 times lower titers than conventional naked DNA vaccines in a P-gal expressing tumor model (132). Vaccination with replicons has also induced protective immunity to melanoma challenge in a TRP-1 expression system, unlike conventional DNA vaccines (133). Although it is

Biological effects of exosomes

In addition to transferring preprocessed antigens in MHC molecules, exosomes might also transfer hsp-associated peptides or cytosolic whole candidate tumor antigens. We could demonstrate that melanoma cells release exosomes containing whole tumor proteins such as MART-1 melanA. Following DC uptake of such melanoma-derived exosomes, cross-presentation of MART-1 peptides by DC MHC class I molecules could be observed. In vivo, DCs pulsed with texosomes induced potent CD8+ T-cell-dependent antitumor effects against established mouse tumors (6). We recently reported that high amounts of tumor exosomes accumulate in the malignant effusions of patients bearing different types of tumors (melanoma, breast, lung, ovarian cancer, mesothelioma) (4). Exosomes harvested from ascitis of melanoma patients efficiently transport MART-1 tumor antigen to monocyte-derived dendritic cells (MD-DCs) for cross-presentation to MART-1-specific CTL clones. Moreover, in in vitro stimulation assays aimed at...

Nucleotide Excision Repair

Marked sensitivity to sunlight (ultraviolet) with subsequent formation of multiple skin cancers and premature death. The risk of developing skin cancer is increased 1000- to 2000-fold. The inherited defect seems to involve the repair of damaged DNA, particularly thymine dimers. Cells cultured from patients with xero-derma pigmentosum exhibit low activity for the nu-cleotide excision-repair process. Seven complementation groups have been identified using hybrid cell analyses, so at least seven gene products (XPA-XPG) are involved. Two of these (XPA and XPC) are involved in recognition and excision. XPB and XPD are helicases and, interestingly, are subunits of the transcription factor TFIIH (see Chapter 37).

Dongchul Kang Zaozhong Su Habib Boukerche and Paul B Fisher

Expression occurring during induction of terminal differentiation in human melanoma cells treated with interferon-beta (IFN-P) plus mezerein (MEZ) (Figure 5). In RaSH, cDNA samples are restriction-digested with frequent cutters such as DpnII to an average size of 256 bp to improve hybridization efficiency. Moreover, restriction digestion of tester cDNA only prior to hybridization enables selection and cloning of tester-unique cDNAs by direct ligation of hybridization mixtures to corresponding sites in the plasmid vector. Thus, RaSH reduces the amount of starting materials required for conventional subtraction hybridization by one tenth by using PCR and obviates the need to separate and clone subtracted cDNA species into plasmid vector or bacteriophage, a nontrivial, technically demanding step in subtractive cDNA library construction (Jiang and Fisher, 1993 Sagerstrom et al., 1997). In addition, RaSH uses reverse Northern Blot hybridization for further rapid confirmation of...

Lymphokine Activated Killer LAK Cells

LAK cells are derived from peripheral blood lymphocytes cultured in the presence of high concentrations of IL-2 for 3-4 d. They demonstrate preferential lysis of tumor cells but not normal cells. Promising data in animal tumor models and preclinical in vitro testing raised the prospect of a readily available source of autologous cells with broad reactivity (12,53,54). Analysis of the relevant precursors of LAK cells indicated a natural killer (NK) origin and the treatment effect was also independent of host T cells (55-57). The first clinical studies of LAK cells were performed at the National Cancer Institute (NCI) surgery branch and used total cell doses ranging from 1.8 to 18.4 x 1010 cells combined with bolus high-dose IL-2 (720,000 U kg). This study showed the greatest efficacy for patients with malignant melanoma or renal cell carcinoma with a 44 overall response rate (58). Based on this promising initial study, several phase II clinical trials of LAK cells were performed in the...

Peripheral Blood Lymphocytes PBLs

Antigen-experienced T cells with a memory phenotype preferentially circulate in the peripheral blood. One of the theoretical advantages of using PBLs is that they are readily accessible and can be safely harvested in large quantities through standard leukapheresis procedures. Several studies tested the clinical efficacy of PBL activated with anti-CD3 and IL-2 termed autolymphocyte therapy (ALT) in patients with metastatic RCC (72,73) or metastatic melanoma (74). Simultaneous CD3 and CD28 stimulation by mAb coupled to magnetic beads in the presence of IL-2 (100-400 IU mL) has also been tested on PBL from cancer patients resulting in 10- to 15-fold numerical expansion after 14 d of culture (75). The PBL stimulated with anti-CD3 alone had equivalent to slightly improved effector function in cytolytic and cytokine release assays compared with combined anti-CD3 anti-CD28 stimulation. Other studies also raise the issue of whether costimulation through CD28 is required to generate superior...

Targets for Antiangiogenic Antilymphangiogenic Gene Therapy 321 Inhibition of Proangiogenic Lymphangiogenic Growth

Simultaneously targeting VEGF production with antisense oligonucleotide and VEGF receptor signaling with receptor tyrosine kinase inhibitors enhances the anticancer efficacy of either therapy alone (96). Some focus has been given to the preclinical gene therapy with soluble truncated forms of VEGFR-2 (sVEGFR-2). This molecule functions in two ways by sequestering VEGF and, in a dominant-negative fashion, by forming inactive heterodimers with membrane-spanning VEGFRs (91,92). Recently, murine fibrosarcoma and melanoma cells transduced with a retrovirus expressing sVEGFR-2, showed tumor growth reduction by inhibition of angiogenesis however, it did not show

Systemic and Local Suppression of T Cells

The indirect influence of tumors on the development of a systemic immunosuppresion can be attributed to hyperproduction of IL-10, oxygen metabolism intermediates, and some enzymes. IL-10 is a type 2 cytokine that is produced by APCs and Th2 cells. It is involved in the development of T-cell anergy, promotion of Th2 responses, and inhibition of Th1 responses, which are important for the generation of efficient antitumor responses. Increased production of oxygen metabolites by macrophages isolated from the metastatic lymph nodes of patients with malignant melanoma was found to be responsible for decreased CD3-mediated stimulation of T cells and the reduction of CTL and NK cell activity (22). The inhibitory effect of macrophages on melanoma-specific CTL lines and NK cells was abrogated in the presence of catalase, a scavenger for H2O2. The mechanism of H2O2-induced immunosuppression by monocytes macrophages derived from the blood of cancer patients was attributed to the inhibition of Th1...

Tumor Targeting 411 Transcriptional Targeting

Transcriptional targeting can be achieved by the use of an expression cassette which is activated by tissue specific promoters (TSPs) (167). The options are to use either a promoter with a high activity in tumor cells tumor ECs or to use inducible promoters to achieve therapeutic transgene expression. Examples of promoters with high tumor-selective activity (minimal expression in normal cells) are, CXCR4, cyclooxygenase-2 (COX-2), survivin (a member of the inhibitor of apoptosis protein family) and pre-proendothelin-1 (PPE-1) a precursor protein for endothelin-1. The human CXCR4 gene is expressed at high levels in many types of cancers, but is repressed in the liver. Thus, the CXCR4 promoter has a tumor-on and liver-off status in vitro and in vivo, which make it a good candidate TSP for targeted cancer gene therapy approaches, (i.e., for melanoma and breast cancers) (168). COX-2, a key enzyme in the synthesis of prostaglandins and thromboxanes, is highly up-regulated in tumor cells,...

Roberta P Glick Terry Lichtor Henry Lin and Edward P Cohen

The prognosis for patients with malignant glioma is poor. Conventional treatments such as surgery, radiation therapy, and chemotherapy have done little to affect long-term survival, and new methods of treatment are urgently needed. In this report, approaches involving cytokine gene therapy in treatment of malignant brain tumors are reviewed and contrasted with a strategy developed in this laboratory involving the use of allogeneic cells which have been genetically modified to secrete cytokines. In our studies, mice with an intracerebral glioma, melanoma, or breast carcinoma treated solely by intratumoral injections with allogeneic cells genetically modified to secrete interleukin-2 were found to survive significantly longer than mice in various control groups. The anti-tumor response was mediated predominantly by T-cell subsets (CD8+ and NK LAK cells). The injections resulted in the killing of only the neoplastic cells non-neoplastic cells were unaffected. Experiments involving...

Association of Specific HLA Class I Molecules With Response to Melacine

If cytolytic T cells were important for achieving a clinical response, we speculated that responding patients might have an overrepresentation of certain specific HLA class I molecules known to restrict CTL responses (20). Seventy patients with metastatic melanoma were studied first. Those patients who had HLA-A2, or the closely related HLA-A28 (now called HLA-A68), HLA-B12 split (including HLA-B44 and HLA-B45) or HLA-C3 had a far greater likelihood of responding to Melacine than those who lacked those alleles. Patients with all three alleles had a 40 response rate, whereas those who lacked all three had nearly 0 response. The group as a whole had a 20 response rate. In a confirmation and extension of these data, Sosman and colleagues (21), analyzing a Southwest Oncology Group study of 689 patients with resected stage II melanoma, found that those with HLA-A2 or HLA-C3 had a longer disease-free survival than those with other HLA phenotypes. Among patients who matched at least two of...

Obtaining Family History Information

Additionally, the pedigree should include information on all surgical procedures in affected and unaffected family members such as bilateral oophorectomy performed either for prophylaxis or for a benign condition or removal of skin lesions. Such surgeries may impact that individuals risk for cancer (as would be the case with oophorectomy) or may indicate another possible unsuspected cancer in the pedigree, such as a melanoma. An example of a cancer pedigree is presented in Figure 1 and important details of the family history are listed in Figure 2.

Other clinical trials with allogeneic lysates

Other investigators have also concluded that allogeneic materials are potentially of greater benefit to patients than autologous tumor-derived immunogens. Among the earliest of these studies were those of Cassel and associates, who utilized viral oncolysates of melanoma cell lines as immunogens (24). These studies were extremely promising, with a number of remissions lasting a decade or more, but unfortunately may have been ahead of their time. Attempts to reproduce the results in large-scale cooperative trials were unsuccessful, but those were hampered by failure of attention to important details, such as viability of the cells before viral treatment. Wallack immunized patients with allogeneic viral oncolysates created with vaccinia virus, and did one of the first double-blind, randomized, multicenter studies in 250 patients perhaps too small a number to show a small difference between the groups with resected stage III melanoma. Protein representing 5 million melanoma cells in...

Conclusions and future

Our personal experience with allogeneic melanoma lysate vaccines, given to several hundred patients, has convinced us that although the response rates in disseminated disease are at most in the range of 15-20 , there is indeed activity of these preparations even when billions of tumor cells are present. There have been a number of patients with a gratifying dramatic increase in survival lasting 5-10 yr or more. Allogeneic lysates may perhaps be effective in preventing recurrence of melanoma in patients with resected stage III disease, and have been equivalent to toxic four-drug chemotherapy even when given by a suboptimal route of administration in a large multicenter trial. With the same caveat, suboptimal (im) route of administration, a large Southwest Oncology Group phase II study of Melacine vs observation in resected intermediate thickness stage II melanoma was initially positive at a follow-up of 1 yr (p 0.04), although further follow-up showed no difference. Melacine was...

Production and purification of exosomes

II concentration of the starting clarified supernatant. This methodology was extended to a miniscale process with comparable results. Likewise, the classical sedimentation technique is a tedious and less productive process carrying over along with exosomes contaminants of the culture medium. Furthermore, the development of quality control assays allowed qualitative and quantitative standardization of exosome preparations. Immunocapture assays assessing exosomal contents in MHC class I and II molecules as well as FACS determination of exosomal protein patterns after coupling onto macroscopic beads are currently used to calibrate exosome dosages in the first phase I trial (24). Exosomes are secreted from monocyte-differentiating DCs from Day 5 to Day 6 using culture conditions devoid of maturing agents (granulocyte-macrophage colony-stimulating factor GM-CSF interleukin-4 IL-4 ). The amounts of exosomal MHC class I molecules recovered in a 24-h DC culture supernatant using such a...

Of In Vivo Activities

Treatment of highly accessible tumors for biopsy will definitely increase our knowledge of the in vivo activity of these genes. There is little technical difficulty in obtaining biopsies from superficial tumors, such as head and neck cancers, as well as melanoma. For tumors which post-treatment biopsy is prevented by technical and ethical reasons (e.g., brain tumors), applying gene therapy as a neoadjuvant therapy (i.e., given before operation) will allow us to analyze the biological endpoints from the resected tumors.

Kelly K Hunt md and Rajagopal Ramesh PhD

The protein encoded by the melanoma differentiation-association gene 7 (MDA-7 IL-24) is a novel interleukin (IL)-10 family cytokine with unique tumor-specific apoptotic and antiangiogenic properties that make it especially attractive for use in cancer gene therapy applications. Mda-7 gene transfer with a replication incompetent adenoviral vector (Ad-mda7) induces apoptosis in a tumor specific manner, an effect that is independent of the status of other tumor suppressor genes, such as p53, Rb, or p16INK4. In addition to its direct cytotoxic effects, Ad-mda7 transduction causes secretion of a processed, glycosylated form of MDA-7 protein. MDA-7 is a novel interleukin (IL-24) with unique apoptotic functions. Studies on the secreted MDA-7 IL-24 protein have shown that it can act as a pro-Th1 cytokine, and induces secretion of interferon-gamma, tumor necrosis factor-a, IL-6, IL-12, and granulocyte macrophage colony-stimulating factor in human peripheral blood mononuclear cells . Additional...

Phase i clinical trials

Tumor vaccines are often based on DNA constructs, viral vectors, and cytokines that have been determined as safe from previous clinical trials. Peptide vaccines generally seem inherently safe so long as the cytokine adjuvants are used in combinations and doses previously demonstrated to be safe. For example, peptide vaccines based on nonmutated melanoma antigens such as MART-1 Melan A and gp100 were initially evaluated in a phase I trial at the National Cancer Institute at doses ranging from 0.1 to 10 mg. However, no toxicity was encountered even at the highest dose (1,2).

Initial Identification and Characterization

The melanoma differentiation associated gene-7 (mda-7) was identified in HO-1 melanoma cells induced to terminally differentiate by treatment with fibroblast interferon (IFN- ), and the protein kinase C activator mezerein (MEZ) (1). The differentiated and growth arrested HO-1 melanoma cells mRNAs were used to generate a cDNA library another library was generated from proliferating HO-1 cells. Differentiation induction subtraction hybridization (DISH) of these two yielded a temporally spaced subtracted cDNA library enriched for genes activated during HO-1 terminal differentiation (2,3). The underlying hypothesis of the above approach was that cancer cells would stop or significantly reduce expression of genes regulating growth control or differentiation, and that treatment with IFN- and MEZ would reactivate them. mda-7 was identified as a gene with minimal or absent expression in proliferating melanoma cells, high expression in normal melanocytes, and inducible expression in terminally...

MDA7is an IL10 Family Cytokine

Chada S. et al. reported that exposure of melanoma cells to MDA-7 IL-24, induced secretion of interferon (IFN)-y and IL-6, but not of IL-4 or IL-5 in contrast, Ad-luc treatment did not induce IFN-y or IL-6, suggesting an MDA-7-specific effect (15). Transduction of these cells with Ad-mda7 induced increases in mRNA that mirrored the cytokine induction observed with exposure to the MDA-7 IL-24 protein. The report suggested that the effect was also specific to some types of tumor cells, because similar treatment of lung and breast cancer cells did not induce release of cytokines. Microarray analysis of non-small-cell lung carcinoma (NSCLC) cells transduced with Ad-mda7 corroborated the cytokine-like activity of this gene, and confirmed that MDA-7 can activate IFN-y and NF-kB signaling pathways. Given that IL-10 functions as an immunosupressive cytokine, and that it significantly inhibited MDA-7 IL-24 activity in human peripherral blood mononuclear cells (PBMCs), it is possible that mda7...

MDA7 is a Tumor Selective Apoptosis Inducing Factor

The initial observation of mda-7 loss of expression in melanomas and its correlation with progression of this type of tumors suggested growth suppressive properties in melanoma cells (4-7). Ensuing studies investigated the effects of ectopic expression of mda-7 in a wide variety of tumor cells (melanoma, carcinomas of the breast, colon, prostate, nasopharynx, high grade gliomas, and osteosarcoma) and proved that mda-7 inhibits tumor cell growth regardless of the status of other genes (p53, Rb, Bax or p16) (1,9,10) (see Fig. 2 for a representative study of lung cancer cells). However, expression of the gene in normal human skin fibroblasts and mammary breast epithelial cells did not significantly affect their growth or trigger apoptosis (9,10). Together, these reports indicate that MDA-7 is an IL-10 family cytokine with tumor cell apoptotic activity and that the cytotoxic effects it induces are specific to tumor cells (5,10,22-24). Several studies have investigated the signal...

Time to Tumor Progression End Point

To detect a large effect of a treatment in delaying tumor progression in a rapidly progressive disease such as pancreatic cancer or melanoma with visceral metastases does not require many patients in a randomized trial. With exponentially distributed times to progression, a 40 reduction in the hazard of progression corresponds to a 67 increase in median time to progression. In order to have 80 power (P 0.20) for detecting this size of effect using a traditional a 0.05, only about 117 patients are required (assuming accrual rate of about 3 patients per month, median time to progression of12 mo for control group and follow-up time of 24 mo after end of accrual) (12). This can be reduced to 87 patients if a 0.10. Hence, 44 patients randomized to vaccine and the same number randomized to control, one can conduct a randomized phase 2.5 trial for evaluating whether the vaccine reduces the hazard of progression by 40 . This design would be a phase 2.5 design because of the unconventional use...

Intraarterial Administration With or Without Blood Brain Barrier Disruption

As shown in experimental animal studies (30,31,134-136), enhancing the delivery of BPA and BSH can have a dramatic effect both on increasing tumor boron uptake and the efficacy of BNCT. This has been demonstrated in the F98 rat glioma model where intracarotid (ic) injection of either BPA or BSH doubled the tumor boron uptake compared to that obtained by iv injection (30). This was increased fourfold by disrupting the BBB by infusing a hyperosmotic (25 ) solution of mannitol via the internal carotid artery. MST of animals that received either BPA or BSH ic with BBB-D were increased 295 and 117 , respectively, compared with irradiated controls (30). The best survival data were obtained using both BPA and BSH in combination, administered by ic injection with BBB-D. The MST was 140 d with a cure rate of 25 , compared with 41 d following iv injection with no longterm surviving animals (31). Similar data have been obtained using a rat model for melanoma metastatic to the brain. BPA was...

Direct Inhibitory Effects on Angiogenesis

Evidence for direct effect of MDA-7-mediated antiangiogenic activity in vivo was next examined. Subcutaneous implantation of MDA-7 producing 293 cells (293-mda7) mixed with A549 lung tumor cells (1 1 ratio) in nude mice resulted in significant suppression of tumor growth compared with tumor growth in mice implanted with a mixture of parental 293 cells and tumor cells (see Fig. 6). That the tumor growth inhibitory effects resulted from exogenous MDA-7 was demonstrated by detecting MDA-7 protein in the tumors. Note that A549 tumor cells do not express detectable endogenous MDA-7 protein. Tumor growth inhibition was demonstrated to occur via apoptotic death of tumor endothelial cells. Associated with tumor growth inhibition was a marked reduction in tumor vascular-ization as demonstrated by the reduced hemoglobin content, and less CD31+ endothelial cells (61). These results demonstrated the direct antiangiogenic activity for MDA-7 IL-24 in vivo. Although these experiments established the...

Sources of Lymphocytes for In Vitro Immune Analysis

The most appropriate source of T lymphocytes for immune analysis is the subject of considerable debate. Clearly, peripheral blood is the most convenient source of T cells to sample, but it is also possible that peripheral blood T-cell activity may not represent the true effector population following immunization. It has furthermore been suggested that peripheral blood T-cell activity may not correlate with clinical response (15). Using human leukocyte antigen (HLA) peptide tetramers, Lee and colleagues (15) enumerated melanoma antigen-specific T-cell precursor frequency directly in peripheral blood mononuclear cells (PBMCs) from melanoma patients vaccinated with gp100 peptide with or without IL-2. Although no antigen-specific T cells could be cultured ex vivo from PBMCs of IL-2-treated patients, these were the only individuals in whom tumor regressions occurred.

Antimetastatic Activity Of Mda7il24

Melanocytes, and mda-7 induction in human hematopoietic cells after treatment with TPA. As mentioned above, mda-7 was initially inversely correlated with melanoma progression (10,22) which is consistent with our results demonstrating that MDA-7 protein expression localizes to superficial areas of primary cutaneous melanoma and decreases as one moves toward the deeper, more invasive, areas of the tumor (6,7). Taken together, these results strongly suggest that MDA-7 expression is lost during melanoma progression and invasion. MDA-7 has reportedly high levels of expression in melanocytes, and in early stage melanomas. Immunohistochemical (IHC) analysis done by our group on a group of paired metastatic and primary melanomas from patients confirms MDA-7 expression at variable levels, in approx 70 of primary tumors. MDA-7 expression significantly decreased in both percent of positive cells and intensity of expression at the deep invasive front of the tumor as compared with its...

Analysis of TCell Receptor V Region Gene Usage and Complementarity Determining Region 3 Sequences

It has been suggested, for some antigens, that antigen-specific T cells have a restricted TCR repertoire (20) that can be detected by sequencing the third complementarity-determining region (CDR3) of the TCR. The CDR3 region encodes the highly polymorphic portion of the TCR responsible for recognizing peptide-MHC complexes. For the P chain, the CDR3 region encodes the Variable (V) region-Diversity (D) segment and Diversity segment-Joining (J) segment junctions, whereas for the alpha chain it encodes the V-J junction. Using V, D, or J-region subfamily specific polymerase chain reaction (PCR) primers, PCR may be performed to detect the development of restricted TCR gene usage (21,22). There is compelling evidence for restricted TCR V-region usage in the response to viral diseases (23,24). Some studies in melanoma patients (25,26) and renal cell carcinoma patients (27) have detected a restricted TCR gene usage. However, other studies in melanoma have found unrestricted TCR gene usage...

Role of cellular immunity in the control of tumor growth

Several studies suggest a beneficial role of proliferating T lymphocytes induced in vaccinated patients with melanoma (1), non-Hodgkin's lymphoma (2), and prostate carcinoma (3-9) (Table 1). These lymphocytes may have provided helper function and, therefore, may have induced cytolytic T lymphocytes (CTLs) that lysed the tumor cells. A positive correlation between CTL induction by vaccination and tumor regression and or enhanced survival has been demonstrated in colorectal carcinoma (CRC) (5), melanoma (6), and non-Hodgkin's lymphoma (9,10) patients. Furthermore, the induction of DTH reactions in vaccinated melanoma patients has been associated with beneficial clinical outcome (7,11-14). These studies emphasize the importance of cellular immunity induction by cancer vaccines for induction of tumor regression and survival enhancement. 1. Major histocompatibility complex (MHC) downregulation or mutation on tumor cells. Downregulation and or mutation of human leukocyte antigen (HLA) class...

Exosomes As Immune System Regulators

Immunization efficiency with exosomes derived from dendritic cells, has began human evaluation (Escudier et al., 2005 Morse et al., 2005) and although a decrease in tumor has been observed owe to antitumoral response induction, is clear that obtaining exosomes from autologous DC is a complicated and impractical process. In contrast, the use of exosomes derived from tumor cells has proof to be useful choice to induce an effective antitumoral response in melanoma and murine plasmocitoma (Altieri et al., 2004 and personal observations). It might be a good alternative then for antitumoral induction if selective in vivo secretion of tumoral exosomes may be induced. In fact, exosomes derived from heat stress tumor cells contains higher concentrations of HSP70, induce a better priming of cytotoxic T lymphocytes (Dai et al., 2005), suggesting that presence of HSP70 is not only important for transport of immunogenic peptides through exosome structures but also during death of tumoral cells....

Association Between Antibody Induction and Tumor Growth Inhibition Increased Survival in Vaccinated Cancer Patients

Several phase I and II clinical trials have demonstrated an association between antibody induction in vaccinated cancer patients and clinical response (enhanced survival, tumor shrinkage) (Table 1). Sialyl-Tn-KLH vaccine induced IgG and IgM antibodies to the antigen in all vaccinated breast cancer patients, and antibody induction was associated with patients' survival (51). Anti-idiotypic antibodies binding to the antigen-combining site of antitumor antibodies may mimic tumor antigen. Anti-idiotypic antibody vaccine mimicking the high-molecular-weight melanoma-associated antigen (HMW-MAA) elicited anti-anti-idiotypic antibodies (Ab3 antibodies binding to anti-idiotype or Ab2) in melanoma patients, and the Ab3 responses were associated with patients' survival (52). However, in neither of the two studies were the antibodies shown to be associated with clinical responses demonstrated to bind to tumor cell-surface determinants, and, therefore, the role of these antibodies in the...

Interstitial Chemotherapy

Recent research by Ewend and co-workers has investigated the use of chemotherapy-loaded wafers as a treatment method for MBT 93-95 . In a mouse model, 20 per cent BCNU-loaded wafers were implanted after surgical resection of melanoma MBT and compared to animals that received only external beam irradiation 93 . Survival was significantly longer in the chemotherapy wafer group (p 0.032), with 57 per cent of the animals still alive at 150 days follow-up. For the radiation alone group, only 50 per cent of animals were still alive during follow-up after 25 days. In a more extensive study using a mouse model, several different MBT (melanoma, renal cell, colon, lung) and chemotherapy wafer (BCNU, carboplatin, camptothecin) types were studied 94 . Each type of wafer was used against each type of MBT, with or without radiotherapy. The results demonstrated a significant survival benefit for the BCNU-loaded wafer over the carboplatin and camptothecin wafers. In addition, there appeared to be a...

Tumor Vaccine Specific Regulatory Issues

Autologous tumor cells are derived from the patient's own tumor. These cells may be modified in vitro before they are injected back into patients with or without adjuvants. In some cases these tumor cells are modified by chemicals, irradiation, or by photochemical treatment in conjunction with ultraviolet (UV) light exposure. These modifications usually render the tumor cells replication incompetent, but certain modifications may enhance immunogenicity as well. It is hypothesized that these modified tumor cells will boost an immune response against tumor antigens and result in rejection of established tumors or development of immune memory that will not allow tumors to recur in cases where tumors were previously resected. Autologous tumor cells have already been used in a number of clinical trials for melanoma, colorectal cancer, ovarian cancer, and renal cancer with moderate success (1).

Overview And Future Considerations

H., Friedman, A. H., and Seigler, H. F. (1998). Demographics, prognosis, and therapy in 702 patients with brain metastases from malignant melanoma. J Neurosurg 88, 11-20. 17. Schouten, L. J., Rutten, J., Huveneers, H. A. M., and Twijnstra, A. (2002). Incidence of brain metastases in a cohort of patients with carcinoma of the breast, colon, kidney, and lung and melanoma. Cancer 94, 2698-2705. 30. Bafaloukos, D., and Gogas, H. (2004). The treatment of brain metastases in melanoma patients. Cancer Treatment Rev 30, 515-520. 44. Franciosi, V., Cocconi, G., Michiarava, M. et a . (1999). Front-line chemotherapy with cisplatin and etoposide for patients with brain metastases from breast carcinoma, nonsmall lung carcinoma, or melignant melanoma. A prospective study. Cancer 85, 1599-1605. 69. Biasco, G., Pantaleo, M. A., and Casadei, S. (2001). Treatment of brain metastases of malignant melanoma with temozolomide. New Engl J Med 345, 621-622. 70. Agarwala, S. S.,...

Chromosomal translocation in synovial sarcomas

A translocation t(X 18) (pll.2 qll.2) occurs consistently in synovial sarcomas. Several genes are located at the breakpoints in this translocation. They are SYT occurring on chromosome 18, and members of the testis cancer antigen SSX gene family, namely SSX1, SSX2 and SSX4 on the X-chromosome. The SSX genes are normally expressed in the testis and the thyroid. Of the SSX homologues, SSX J, SSX2 and SSX4 are frequently expressed in human neoplasms. SSX3 is not usually detected. Furthermore, SSX expression is found in many forms of cancer including breast cancer, colorectal cancer, head and neck tumours, melanoma and in lymphomas. No expression of the SSX genes is detectable in leukaemias, thyroid cancers, and seminomas (Tureci et al., 1998).

Cellular Delivery Methods

Another delivery method for both viral and non-viral vectors is ex vivo transduction of target cells that are then reintroduced into the patient (6). This delivery mechanism was first used by Rosenberg et al. in the initial human gene therapy trial when human tumor infiltrating cells were transduced with retroviral constructs and reinfused in metastatic melanoma patients (1). The advantage of this delivery method is that there is more control of the transduced cell and higher transduction efficiencies can be achieved in the target tissue. Disadvantages included difficulties in targeting the reinfused cells to specific anatomic locations as well as possible contamination with reintroduced biologic agents. Current research efforts are evaluating ex vivo transduction of stem and progenitor cells which may lead to more sustained gene expression and better tissue targeting.

Stimulating the Immune System

Antigens that are capable of provoking weak humoral or cellular reactions. By activating this immune response against tumor cells through gene transfer it is hoped that tumors can be eradicated either by the transferred gene product or activation of the patient's own immune system. In animal models, the administration of cytokines such as inter-leukin (IL)-2, IL-4, IL-6, IL-7, IL-12, tumor necrosis factor (TNF)-a, interferons and granulocyte macrophage colony-stimulating factor (GM-CSF) have resulted in tumor regression (8). The systemic administration of cytokines in human trials though has been limited by the severe toxicity of these cytokines. Gene therapy strategies offer an opportunity to overcome these limitations because of the potential for local delivery to injected tumor with reduction in systemic toxicity (9-11). Another approach to make tumor cells more immunogenic involves the cotransfer of stimulatory molecules such as B7.1 and B7.2. These molecules provide a key event...

Tumor Suppressor Gene Replacement

Tumor suppressor genes are a class of genes whose absence may contribute to tumor growth. In most situations tumor suppressor genes require both alleles of a gene to be deleted or inactivated to lead to tumor growth. The replacement of just one functional tumor suppressor gene may therefore be enough to restore normal growth regulation and induce tumor apoptosis. The tumor suppressor gene with the most clinical trial experience is the gene encoding for wild type (wt)-p53. The wt-p53 gene may inhibit tumor development by either suppressing genes that contribute to uncontrolled cell growth and proliferation or activating genes that inhibit cell growth. Functional p53 is normally responsible for detecting damaged DNA and either directing repair of damaged cells or committing cells to apoptosis (programmed cell death) if the DNA is not able to be repaired. Several wt-p53 gene replacement strategies are in clinical trials for head and neck cancer, melanoma, breast, brain, and lung cancer...

Adenovirus use in cancer vaccine strategies

Another type of cancer presenting attractive targets for cancer vaccination approaches is melanoma. The melanosomal proteins TRP2, MART, and gp100 are all highly expressed in melanoma, and have been studied in experiments involving recombinant Ad vaccines. TRP2 is highly homologous in mice and humans, and although attempts at plasmid and gene gun vaccination have failed to induce immunity, administration of an Ad vector expressing a xenogenic human TRP2 caused mice to mount an immune response against their native melanocytes, resulting in coat depigmentation (23). These mice were also significantly protected against metastatic growth of B16 melanoma, an immunity associated with the presence of TRP2-reactive antibodies and CD8 T cells. In vitro studies of HLA-A2+ cell lines confirmed this association, in showing generation of melanoma-specific CD8 T cells for the other melanoma antigens, gp100 and MART-1 (24). Studies conducted in C57Bl 6 mice mirror results found with TRP2, showing a...

Analytic Epidemiology

An insight into potential risk factors for HS may come from analyses of disease associations since associated diseases may share common risk factors. In principle, associations may also result from exposures which follow the development of one of the diseases of interest (e.g., iat-rogenic factors) or even represent an artifactual effect if the presence of a concomitant disease influences the diagnosis of another disease or its referral (Berkson's bias). Interestingly, in a large-scale analysis of Swedish hospital discharge diagnoses linked with data from the Swedish National Cancer Registry, a strong association was documented between HS and non-melanoma skin cancer (NMSC), buccal cancer and liver cancer 16 . These associations may at least partly reflect chance findings derived from multiple testing. However, the primary hypothesis of the study was centered on an association between HS and NMSC. Even if NMSCs complicating perineal or buttock HS localizations is a possible...

Patients With Cancer Can Be Immunized With Class II Peptide Based Vaccines

A successful vaccine strategy for generating peptide-specific CTLs capable of lysing tumor expressing HER-2 neu and resulting in durable immunity involved immunizing patients with putative Th epitopes of HER-2 neu that had, embedded in the natural sequence, HLA-A2-binding motifs of HER-2 neu. Thus, both CD4+ T-cell help and CD8+-specific epitopes were encompassed in the same vaccine. In this trial, 19 HLA-A2 patients with HER-2 neu-overexpressing cancers received a vaccine preparation consisting of putative HER-2 neu helper peptides (40). Contained within these sequences were the HLA-A2-binding motifs. Patients developed both HER-2 neu-specific CD4+ and CD8+ T-cell responses. The level of HER-2 neu immunity was similar to viral and tetanus immunity. In addition, the peptide-specific T cells were able to lyse tumor. The responses were long-lived and detectable for greater than 1 yr after the final vaccination in selected patients. These results demonstrate that HER-2 neu MHC class II...

Cadher1ns And Intercellular Adhesion

Integrin Cadherin

Catenins are now known to be key regulatory molecules that mediate the transduction of extracellular contacts between cadherins during epithelial reorganization and also provide for the linkage of cadherins to intracellular signaling pathways reviewed in 118-120 . Importantly, catenins transmit signals that regulate gene expression. Thus, p-catenin binds both cadherin and other catenins, but when P-catenin is in excess, it binds the LEF family of transcription factors, and this complex of catenin and transcription factor is transported to the nucleus where it effects gene expression. In contrast, expression of the wnt-1 protooncogene increases the level and function of catenins, thereby stabilizing cadherin-mediated cell-cell adhesion. Wnt signaling involves GSK3-P, and through the phosphorylation of P-catenin, its own turnover is regulated. Also, axin binds p-catenin, APC, and GSK3-p directly, and this tetrameric complex regulates stabilization of P-catenin 121,122 , Finally, pl20ctn...

CMyc Deregulation in Cancer

Myc Deregulation

C-Myc overexpression is associated with neoplasms of different tissues, including breast (11,12), neuroblastoma (13), cervical carcinoma (14), malignant melanoma (15), prostate cancer (16), osteogenic sarcoma (17), and lymphoid cancer (6, 18, 19) (reviewed in 20). Thus, Myc protein overexpression is an important player in cellular transformation (21-25).

Hydroxylation by monophenol monooxygenase

This type of reaction usually forms catechols from phenols. Its physiological importance in vertebrates is the formation of l-dopa from l-tyrosine by tyrosinase, usually in skin l-dopa is the precursor of melanin. The presence of unusually high urinary concentrations of intermediate metabolites associated with melanin formation is observed (in man) both after exposure to sunlight, and in patients with melanoma without exposure to the sun. These metabolites can be used as a marker in the diagnosis of melanoma. In mouse pups devoid of tyrosine hydroxylase tyrosinase appears to be responsible for the appearance of significant amounts of neural catecholamines K89 .

Tyramine 3hydroxylase

Vibrio tyrosinaticus is composed of two tyrosinases, molecular weights 41 000 and 38 500 it does not cross-react with antiserum to hamster melanoma tyrosinase. It acts on l-tyrosine and slightly on the d- isomer and m-tyrosine, but catechol and l-phenylalanine are not substrates. It is inactivated by diethyldithiocarbamate, and this is reversed by Cu2 + , Mn2 + , Cd2+ or Fe2 + A141 .

Chemoprotection And Antimutagenic Effects

An in vitro study on human bronchial cells found that rosemary extract and its constituents, carnosol and carnosic acid, may have chemoprotective activity through decreasing carcinogen activation via inhibition of the enzyme cytochrome P450 (CYP1A1) and increasing carcinogen detoxification by induction of phase II enzymes (Offord et al 1995). Carnosol has been found to also restrict the invasive ability of mouse melanoma cells in vitro by reducing MMP-9 expression and activity (Huang et al 2005).

Cancer In Werner Syndrome

WS patients are at increased risk of developing cancer (Goto et al., 1996 Monnat, 2001 Monnat, 2002). The elevated risk of neoplasia in WS patients is of particular biological interest. As discussed later, neoplasia may be an expression of important mechanistic links between WRN function in vivo, genome stability assurance, and the limitation of cell proliferation defects. The elevated risk of neoplasia in WS is selective in that only a small subset of neoplasms are clearly elevated in incidence as compared with general population controls (see Table 80.2). The following neoplasms, in order of decreasing frequency, have been observed most often in WS patients and occur at higher or much higher frequency than in normal population controls soft tissue sarcomas, thyroid carcinoma, meningioma, malignant melanoma, malignant or preneoplastic hematologic disease, and osteosarcoma. Many other neoplasms, including common adult epithelial malignancies, have been observed in WS patients....

Tools and Targets of Gene Therapy

Muscle Implants For Muscular Dystrophy

Melanoma Another genetic approach to battling cancer is to enable tumor cells to produce immune system biochemi-cals, or to mark them so that the immune system more easily recognizes them. This approach is called a cancer vaccine. A treatment for the skin cancer melanoma, for example, alters tumor cells to display an antigen called HLA-B7, which stimulates the immune system to attack the cell.

Carbohydrate Cell Surface Cancer Antigens The Mskcc Experience

We have screened a variety of malignancies and normal tissues with a series of 40 monoclonal antibodies against 25 antigens that were potential target antigens for immunotherapy (18-21). Results for the 12 defined antigens expressed strongly in 50 or more of biopsy specimens of breast, ovary, prostate cancer, melanoma, sarcoma, and small-cell lung cancer (SCLC) are shown as examples in Table 2. The 13 excluded antigens (including CEA carcinoembryonic antigen and HER2 neu) were expressed in 0-2 of the 5-10 specimens. Melanoma

Richard S Foster Md Richard Bihrle Md John P Donohue Md

Rplnd Anatomy

What is sometimes underappreciated in regard to therapy for testicular cancer is that not only is metastatic testicular cancer chemosensitive, it is also surgery sensitive. The surgical removal of metasta-tic testicular cancer of the retroperitoneum, pelvis, lungs, mediastinum, or visceral organs is curative from 30 to 75 of the time, depending on the site of metastasis and the clinical situation. When one compares this ability to cure with surgical removal alone in cases of testicular cancer with the ability of surgical removal to cure in cases of other cancers (such as breast cancer, colon cancer, melanoma, etc), one can recognize that testicular cancer is unique from a surgical point of view. Furthermore, testicular cancer is amenable to surgical cure not only in patients at a low stage of disease but also in those patients who have completed chemotherapy or are resistant to chemotherapy. Even in the setting of chemorefrac-tory disease localized to the retroperitoneum, cure at the...

TCell Anergy Tolerance

In clinical settings, the circulating CD8+ T cells, specific for melanoma-associated antigens MART-1 or tyrosinase, were found to be unable to lyse melanoma target cells or produce cytokines. The ability of these cells to lyse Epstein-Barr virus-pulsed target cells or generate allogeneic responses was not impaired (31), indicating the specificity of anergy. Stimulation of T cells from melanoma patients with melanoma peptides did not induce the upregulation of CD69 expression, indicating the T-cell functional defects (31). Vaccination of melanoma patients increased frequencies of melanoma antigen-specific CD8+ T cells identified by human leukocyte antigen (HLA) peptide tetramers, however, no tumor regression was determined (32). Other studies demonstrated that vaccination-induced T cells with an increased level of IFN-y mRNA, which, however, was not accompanied by a significant inflammatory response or CD4+ CD8+ T-cell accumulation (33,34). Detection of these antigen-reactive T...

Overview Of Biomarkers

To compare the DNA-repair capacities of basal cell carcinoma (BCC) in skin cancer patients and controls (119). An age-related decline in DNA-repair capacity was detected, and reduced repair capacity was a particularly important risk factor for young individuals with BCC and for those with a family history of skin cancer. Younger individuals with BCC repaired DNA damage poorly when compared with controls. With increasing age, however, differences between cases and controls gradually disappeared (120). The normal decline in DNA repair observed with increasing age may account for the increased risk of skin cancer that begins in middle age, which suggests that the occurrence of skin cancer in the young may represent a biochemical manifestation of decreased repair capacity. The same technique has been applied in another case-control study (121) of 51 newly diagnosed lung cancer patients and 56 age-, sex-, and ethnicity-matched controls. The mean level of DRC in cases (3.3 ) was...

The Core Data in Histopathology Specimens

Other descriptors include unifocality (pT1a) versus multifocality (pT1b), lymphatic invasion (L), venous invasion (V), classification during or after multimodality therapy (ypT), recurrent tumour (rpT) and multiple primary tumours (pTm). Qualifying tumours in the TNM system are carcinoma, malignant mesothelioma, malignant melanoma, gestational trophoblastic tumours, germ cell tumours and retinoblastoma.

Examples Of These Models In

This case highlights the shortcomings of all the models when a family history falls outside the strict remit of the model. The family history depicted in the pedigree is strongly suggestive of a mutation in BRCA2 to the experienced clinician, given the additional prostate cancer and melanoma on a background of early-onset breast cancer and ovarian cancer. Both these types of cancer have been found to be associated with BRCA2 mutations rather than BRCA1 mutations in recent studies EMBRACE, unpublished, Thompson et al. (37) . Out of the models above, only the Manchester scoring system and BOADICEA consider the additional cancers, and neither of them incorporates melanoma into their calculations yet. It is unexpected that the Myriad risk assessment for this family is identical to that for the family in Pedigree 1. This reflects the limitations of that model in terms of the number of relatives it assesses and the cancers it will consider. This pedigree also serves to demonstrate further...