[Impact of СD8/FoxP3 T lymphocyte ratio in the peritumoral area of primary cutaneous melanoma on the prognosis of the disease]. / Vliyanie sootnosheniya CD8/FoxP3 T-limfotsitov v peritumoral'noi zone pervichnoi melanomy kozhi na prognoz techeniya zabolevaniya.

UNLABELLED: Cutaneous melanoma (CM) is a malignant tumor characterized by typical histological features, one of which is tumor-infiltrating lymphocytes (TIL) that reflect the state of local immunity and determines the course of the disease. AIM: to study a correlation of the ratio of CD8/Foxp3 T lymphocyte subpopulations infiltrating primary cutaneous melanoma (PCM) with the clinical and morphological factors and prognosis of the disease. MATERIAL AND METHODS: The CD8+ and FoxP3+ T-lymphocyte subpopulations infiltrating PCM were investigated in 180 cases by immunohistochemical staining with anti-CD8 and anti-FoxP3 antibodies. RESULTS: The predominant type of TIL was CD8+ cytotoxic T lymphocytes (80.1 cells in the field of vision); FoxP3+ T lymphocytes averaged 34.9 cells in the field of vision. There was a statistically significant correlation of a high ratio of CD8/FoxP3 T cells with disease stage, ulceration and regression segments in PCM, a low disease progression rate, and higher 5-year overall and relapse-free survival. CONCLUSION: The findings may be used in researches and in the practical work of pathologists and clinicians.

[ Spectrum of oncogene mutations is different in melanoma subtypes].

Melanoma is the most lethal malignancy of skin, which is comprised of clinically relevant molecular subsets defined by specific "driver" mutations in BRAF, NRAS, and KIT genes. Recently, the better results in melanoma treatment were obtained with the mutation-specific inhibitors that have been developed for clinical use and target only patients with particular tumor genotypes. The aim of the study was to characterize the spectrum of "driver" mutations in melanoma subtypes from 137 patients with skin melanoma and 14 patients with mucosal melanoma. In total 151 melanoma cases, the frequency of BRAF, NRAS, KIT, PDGFRA, and KRAS mutations was 55.0, 10.6, 4.0, 0.7, and 0.7%, respectively. BRAF mutations were found in 69% of cutaneous melanoma without UV exposure and in 43% of cutaneous melanoma with chronic UV exposure (p=0.045), rarely in acral and mucosal melanomas. Most of melanomas containing BRAF mutations, V600E (92%) and V600K (6.0%) were potentially sensitive to inhibitors vemurafenib and dabrafenib. NRAS mutations were more common in cutaneous melanoma with chronic UV exposure (26.0%), in acral and mucosal melanomas; the dominant mutations being Q61R/K/L (87.5%). KIT mutations were found in cutaneous melanoma with chronic UV exposure (8.7%) and mucosal one (28.6%), but not in acral melanoma. Most of KIT mutations were identified in exon 11; these tumors being sensitive to tyrosine kinase inhibitors. This is the first monitoring of BRAF, NRAS, KIT, PDGFRA, and KRAS hotspot mutations in different subtypes of melanoma for Russian population. On the base of data obtained, one can suppose that at the molecular level melanomas are heterogeneous tumors that should be tested for "driver" mutations in the each case for evaluation of the potential sensitivity to target therapy. The obtained results were used for treatment of melanoma patients.

Patient perception of the benefit of a BRAF inhibitor in metastatic melanoma: quality-of-life analyses of the BREAK-3 study comparing dabrafenib with dacarbazine.

BACKGROUND: In a randomized phase III study (BREAK-3), dabrafenib showed prolonged progression-free survival (PFS) (median 5.1 versus 2.7 months; hazard ratio = 0.30; 95% confidence interval 0.18-0.53; P < 0.0001) compared with dacarbazine (DTIC) in patients with BRAF V600E metastatic melanoma. Assessing how these results are transformed into a real health benefit for patients is crucial. METHODS: The EORTC QLQ-C30 questionnaire assessed quality of life (QoL) at baseline and follow-up visits. RESULTS: For DTIC, all functional dimensions except role dimension worsened from baseline at follow-up. For dabrafenib, all functionality dimensions remained stable relative to baseline or improved at week 6; mean change in seven symptom dimensions improved from baseline, with appetite loss, insomnia, nausea and vomiting, and pain showing the greatest improvement. In the DTIC arm, symptom dimensions were unchanged or worsened from baseline for all symptoms except pain (week 6), with the greatest exacerbations observed for fatigue and nausea and vomiting. Mixed-model-repeated measures analyses showed significant (P < 0.05) and/or clinically meaningful improvements from baseline in favor of dabrafenib for emotional and social functioning, nausea and vomiting, appetite loss, diarrhea, fatigue, dyspnea, and insomnia at weeks 6 and/or 12. After crossing over to dabrafenib upon progression (n = 35), improvements in all QoL dimensions were evident after receiving dabrafenib for 6 (n = 31) to 12 (n = 25) weeks. CONCLUSIONS: This first reported QoL analysis for a BRAF inhibitor in metastatic melanoma demonstrates that the high tumor response rates and PFS superiority of dabrafenib over DTIC is not only a theoretical advantage, but also transforms in a rapid functional and symptomatic benefit for the patient. ClinicalTrials.gov Identifier: NCT01227889.

Functional and symptom impact of trametinib versus chemotherapy in BRAF V600E advanced or metastatic melanoma: quality-of-life analyses of the METRIC study.

BACKGROUND: In a randomized phase III study, trametinib prolonged progression-free survival and improved overall survival versus chemotherapy in patients with BRAF V600 mutation-positive melanoma. PATIENTS AND METHODS: Patients' quality of life (QOL) was assessed at baseline and follow-up visits using the European Organisation for Research and Treatment of Cancer Core QOL questionnaire. RESULTS: In the primary efficacy population (BRAF V600E+, no brain metastases) from baseline to weeks 6 and 12, patients' global health status scores worsened by 4-5 points with chemotherapy but improved by 2-3 points with trametinib. Rapid and substantive reductions in QOL functionality (e.g. role functioning, 8-11 points at weeks 6 and 12) and symptom exacerbation (e.g. fatigue, 4-8 points; nausea and vomiting, 5 points, both at weeks 6 and 12) were observed in chemotherapy-treated patients. In contrast, trametinib-treated patients reported small improvements or slight worsening from baseline at week 12, depending on the functional dimension and symptom. The mean symptom-scale scores for chemotherapy-treated patients increased from baseline (symptoms worsened) for seven of eight symptoms at week 6 (except insomnia) and six of eight symptoms at week 12 (except dyspnea and insomnia). In contrast, at weeks 6 and 12, the mean symptom-scale scores for trametinib decreased from baseline (symptoms improved) for pain (11-12 points), insomnia (10-12 points), and appetite loss (1-5 points), whereas those for diarrhea worsened (15-16 points). Mixed-model repeated-measures analyses showed significant (P < 0.05) and/or clinically meaningful improvements (small to moderate) from baseline in favor of trametinib for global health; physical, role, and social functioning; fatigue; pain; insomnia; nausea and vomiting; constipation; dyspnea; and appetite at weeks 6 and/or 12. QOL results for the intent-to-treat population were consistent. CONCLUSIONS: This first QOL assessment for a MEK inhibitor in metastatic melanoma demonstrated that trametinib was associated with less functional impairment, smaller declines in health status, and less exacerbation of symptoms versus chemotherapy.

[Use of sunitinib in metastatic renal cell carcinoma patients with poor prognosis].

We evaluated overall and relapse-free survival in patients with metastatic renal cell carcinoma (MRCC) with poor prognosis. The study involved 92 patients (median age 54.1 years, male 63%, female 29%), retrospectively; 48.9% had poor prognosis. They received oral sunitinib 59 mg/day in 6 cycles (4 weeks on, 2 weeks off). Median overall survival was 17.6 months, relapse-free--10.8 months. Risk of progression in the poor and good prognosis groups was identical (HR=09). Overall response was 29% in poor prognosis and 40%--in good one (p = 0.07). Drug tolerability was acceptable in both groups. Sunitinib was effective in both groups of MRCC patients.

Use of recombinant interleukin-2 for intrapleural therapy of tumor-associated pleurisy.

Clinical efficiency of low dose Roncoleukin was studied in 30 patients with metastatic exudative pleurisy. Intrapleural therapy proved to be highly effective (overall effect reached 84%), was well tolerated, and improved patients' quality of life.

[Cancer vaccinotherapy: from the experiment to clinical application].

Methods of cancer immunotherapy with dendritic cell-based vaccines, activated outside the organism, as well as techniques employing genetically modified whole-cell antitumoral vaccines, transfected by cytokine gens and stimulating the patient's dendritic cell stimulation in vivo, have been developed in Russian Oncological Research Center. Phase 1 clinical trials of dendritic cell vaccine in patients with disseminated melanoma and colon cancer have been carried out. The treatment is well-tolerated; significant adverse effects are absent. Moderate clinical efficacy and immune activation have been noted. Preclinical and phase 1 clinical studies of genetically modified vaccines also demonstrate the safety of this treatment and its ability to activate antitumoral immune response. Both vaccines are promising as subjects of rationally planned clinical trials of the efficacy of this treatment.

[Adoptive immunotherapy of malignancies].

The results of the studies show that mononuclear leukocytes isolated from human peripheral blood, malignant effusions, and surgically removed spleen can be generated into lymphokine-activated killers (LAK) characterized by high anti-tumor activity. LAKs may be used foradoptive immunotherapy of malignant effusions and the prevention of tumor recurrence in oncological patients after radical surgery.

[Melanoma cell lines as the basis for antitumor vaccine preparation].

The aim of the study was to obtain cell lines from tumor samples, and to determine phenotypic cell characteristics in order to choose the optimal line for vaccine preparation. 15 cell lines with stable growth, varying in cultural growth character and cytomorphology, were obtained from samples taken from patients with metastatic skin melanoma. Immunofluorescense method was used to determine the expression of T- and B-lymphocyte markers, antigens of major histocompatibility complex (MHC) class I and II, and CD86 co-stimulating molecule in the cell lines. The expression of melanocyte differentiation antigens and cancer/testicular antigens was evaluated using immunocytochemical assay. The results allowed the authors to distinguish three types of melanoma cell lines according to the expression of MHC molecules: MHC-negative; MHC class I positive; MHC classes I and II positive.

[Search for HHV-8 associated diseases reservoir and spread paths of HHV-8 in Russia]. / Poiski HHV-8-assotsiirovannykh zabolevanii, rezervuara i putei rasprostraneniia HHV-8 v Rossii.

Summarized in the paper are study results of human herpesvirus type 8 (HHV-8) and of its association with Kaposi's sarcoma (KS). The data obtained denotes that the share of individuals producing the antibodies to HHV-8 in a majority of studied patients was low and ranged form 0 to 5.5%, which is indicative of a low degree of the virus spread in population. At the same time, a high share of persons with antibodies to HHV-8 was detected among HIV-infected homosexuals (71.4%), kidney recipients (26.0%) and among AIDS-KS patients (78.6%). It was also unexpectedly high among patients with T- and B-cell lymphomas (50%), encephalopathy (27.3%) and with stomach cancer (41.8%): the appropriate parameters were 7-12-fold higher versus healthy subjects. The HHV-8 markers, i.e. virus specific antibodies and/or nucleotide sequences of the virus, were detected in blood serum and ejaculate of a significant number of patients with different pathologies of the prostate. Such detection of viral markers in the above categories of patients is suggestive of that sexual contacts with such patients are decisive for the HHV-8 spread in population.

[Retrospective evaluation of the safe reduction of surgical margins of resected stage-IIA cutaneous melanoma of the trunk and extremities]. / Retrospektivnaia otsenka bezopasnogo umen'sheniia granits rezektsii dlia melanomy kozhi tulovishcha i konechnostei IIA stadii.

The paper deals with analysis of 102 case histories locally-advanced 1.51-4.00 mm-thick cutaneous melanomas (CM) of the trunk and arms and legs operated on at the Center's Clinics and 52--at the Regional Oncological Dispensary, Samara. The effectiveness of relatively conservative procedures of treating CM of "medium" thickness and "intermediate" prognosis were assessed by histological analysis of resected material. CMs with such characteristics conform to the specifications of stage IIA of the criteria used by the American Joint Committee on Staging of Cancer (AJCC). The study of the time and frequency of relapse and dissemination of tumor in 154 patients provided the guide-lines for determining optimal extent of surgery to excise CM stage IIA of the trunk and arms and legs. Excision of 1.5-4.00 mm-thick CMs with 2 cm-wide margins left should be considered safe and less traumatic.

Chronic administration of interferon-a decreases blood pressure and heart rate in rats.

We studied the effects of interferon-alpha on rat cardiovascular system. Intravenous administration of intron-A in a dose of 100,000 IU/kg for 3 days led to a permanent and statistically significant decrease in blood pressure (on days 2 and 3) and reduction in heart rate. These effects were not associated with changes in baroreflex regulation of the cardiovascular system.

[Surgical treatment of superficial melanoma of the skin]. / K voprosu o khirurgicheskom lechenii poverkhnostnykh form melanom kozhi.

The 12-year end-results of standard "wide" and sparing "narrow" excision of superficial skin melanoma are compared. The results were contributed by a joint study of the WHO Collaborating Centers for Evaluation of Methods of Diagnosis and Treatment of Melanoma headed by Dr. U. Veronesi and Dr. N. Cascinelli and 23 other centers in different countries (the Blokhin and Petrov Centers in Russia). The investigation comprised 612 patients. After randomization, "narrow" excision of primary tumor with 1 cm-wide margins was performed in 305 patients. In the remaining 307 patients, "wide" excision left margins within 3 cm. Tumor was identified as superficial on the basis of thickness (Breslow), the threshold being 2 mm-thick invasion. This same prognostic indicator was used in both groups. Another one was 12 year-long recurrence-free survival. The sparing excision of primary cutaneous melanoma with a thickness under 2 cm proved effective.

[Clinico-genetic aspects of cutaneous melanoma. I. Incidence, familial study, genetic heterogeneity]. / Kliniko-geneticheskie aspekty melanomy kozhi. I. Rasprostranennost', semeinoe izuchenie, geneticheskaia geterogennost'.

Epidemiologic and clinical genetic data on cutaneous malignant melanoma (CMM) are presented. The incidence of CMM in Moscow from 1983 to 1987 was analyzed. Cumulative incidence (cumulative risk) was calculated for various life periods on the basis of estimates for various age groups, which were used as population frequencies. By the age of 85, these frequencies were 0.35% for males and 0.38% for females. Among relatives of patients, the incidence of CMM was 1.420 +/- 0.498% for males and 1.110 +/- 0.348% for females; i.e., the incidence exceeded the population frequency by a factor of 3 or 4. As a whole, the familial frequency of cancer was equal to 13.3% for male probands and 14.2% for female probands, i.e., more than three times higher than the population frequency. The data obtained formed the basis for the development of CMM classification. hereditary and nonhereditary variants of cutaneous melanoma were identified. Familial cases and CMM that occurred against the background of inherited disease or syndromes were classified as hereditary variants of CMM; taken together, they accounted for 38.8%. The data provided grounds for identification of families at increased risk for the development of CMM.

[Clinico-genetic aspects of cutaneous melanoma. II. Interconnection and pathogenetic commonality with dysplastic nevus syndrome]. / Kliniko-geneticheskie aspekty melanomy kozhi. II. Vzaimosviaz' i patogeneticheskaia obshchnost' melanomy kozhi s displasticheskimi nevusami.

Evidence for the role of dysplastic nevi (DN) in the development of cutaneous malignant melanoma (CMM) is presented. Primary multiple foci of CMM were found considerably more frequently in individuals with DN. The frequency of primary multiple CMM was found to be 3.1% in males with DN and 0.9% in males without DN; in females, 6.8 and 0.6%, respectively. Genetic correlation analysis was performed to determine the genetic interrelation between DN and CMM. In general, the genetic correlation coefficient was 0.9; i.e., predisposition to DN and CMM is determined by common genes for 90%. The frequency and distribution of constitutive fragile sites in chromosomes of peripheral lymphocytes was studied by the method of principal components for discrete variables. The site 1p22 is responsible for variability of the traits CMM and DN for 98.5%. On the one hand, this suggests that one of the supposed genes for CMM can be located at 1p22; on the other hand, CMM and DN are likely to have a common genetic determination or to be very tightly linked. Estimates of risk for the development of CMM in patients' relatives are given with reference to the variants of CMM manifestation and presence of DN.

[Lysosomal proteolytic enzymes in the processes of melanoma invasion and metastasis]. / Lizosomal'nye proteoliticheskie fermenty v protsessakh invazii i metastazirovaniia melanom.

Spectrofluorimetry was used to study the activity of lysosomal and membrane-binding endopeptidases such as cathepsins B (CB), D (CD) and L (CL) in 72 patients with skin malignant melanoma. Lysosomal endopeptidases were measured separately in the primary tumor, normal skin adjacent to tumor and intact and metastatic lymph nodes. The activity of lysosomal CB in the primary tumor was higher than in the adjacent normal skin. The peak activity of membrane-binding KB was established in metastatic lymph nodes. A significant increase in the activity of lysosomal KB was observed in involved regional lymph nodes as compared to intact ones. The activity of membrane-binding KB in metastatic lymph nodes was 6 times that in metastasis-free ones. A significant increase in the activity of lysosomal KD was established in primary melanoma nodes compared to adjacent normal skin as well as in metastatic lymph nodes compared to intact ones. Preoperative chemo- and radiation therapy was followed by a decrease in lysosomal KB and KL activity in metastatic lymph nodes which, however, did not reach the level established for intact lymph nodes. The pathogenetic role of proteolytic endopeptidases in invasion and dissemination of malignant melanoma is discussed as well as the value of their level measurement for assessing metastatic potential of tumor and prognosis of disease on the basis of tumor site, degree of invasion regional lymph node status.