bluuebunny/crossref_metadata_2025_split huggingface dataset:
Early alterations of microRNA expression to predict and modulate melanoma metastasis.
Authors: Hernando Eva , Hanniford Douglas , Shang Shulian , Segura Miguel F. , Pavlick Anna C. , Berman Russell S. , Shapiro Richard L. , Darvishian Farbod , Osman Iman , Shao Yongzhao 
Published: March 2020

8550 Background: Melanoma is curable for most patients whose tumors are surgically removed early in disease progression; however, many primary melanomas recur and progress to metastasis. Clinical staging is insufficient to robustly classify patients at highest risk of recurrence, and prognostic molecular biomarkers have not yet been identified. Methods: We performed miRNA profiling of 92 FFPE primary melanomas to discover metastasis relevant miRNAs and develop predictive models of recurrence, which were then validated in an independent cohort. We identified miRNAs differentially expressed between primary tumors that did and did not recur (3-year minimum follow-up) and between thick and thin lesions. We selected candidate miRs for screening in a fluorescence-based in vitro invasion assay, and prioritized a subset for in vivo testing. Potential downstream mediators of these miRNAs were selected by mRNA array analysis and tested in a secondary invasion screen. mRNA candidates that mimicked the miR’s invasion-suppressive effect were tested in 3’UTR reporter assays to confirm them as direct targets. Results: Using the discovery cohort we identified a 20 miRNA signature that can distinguish Stage I/II primary tumors (n=70) that did from those did not recur with 3-year minimum follow-up with an AUC=94%, 95% CI: (0.88, 0.99). Applying this model to predict risk for recurrence in the independent validation cohort yielded an AUC = 96%, 95% CI: (0.90, 1) in discriminating recurrent versus non-recurrent stages I/ II patients (n=45). From the discovery cohort, 40 candidates were selected for invasion assay screening, of which 5 miRNAs robustly inhibit in vitro invasion in 5 melanoma cell lines. Three miRs (miR-382, miR-516b, and miR-7) strongly suppressed metastasis in a mouse model. Moreover, multiple mRNAs tested as potential mediators mimicked the invasion-suppressive effects of candidate miRs. Of those, NCAPG2 and CDC42 were identified as miR-516b targets, CTTN as a miR-382 target, and PIK3CD as a miR-7 target. These genes have been linked to metastasis in melanoma or other tumors. Conclusions: Our data demonstrate that aberrant expression of specific miRNAs at diagnosis is predictive of and functionally impacts melanoma progression.
Analysis of KIT expression and mutations in sinonasal, genital, and acrolentiginous melanomas.
Authors: Bull Christian R F , Schoenewolf Nicola L , Belloni Benedetta , Holzmann David , Tonolla Sabina , Lang Roland , Mihic Daniela , Andres Christian , Dummer Reinhard 
Published: March 2020

8563 Background: Melanoma is subdivided into molecular defined groups. One major subtype is characterized by a mutation in the BRAF gene. BRAF is less commonly mutated in acrolentiginous (ALM) and mucosal (MM) melanomas, instead these subtypes predominantly show aberrations in the KIT/CD117 gene. KIT aberrations predict the outcome of targeted therapies in ALM and MM patients, however the particular role of KIT expression in these melanoma subtypes remains controversial. To explore the relationship between KIT expression, mutation, and tumor stages, we investigated immunohistochemical KIT expression and mutation status in primary ALM/MM lesions and their metastases. We also compared the frequency of KIT mutations of MM in different anatomic locations. Methods: KIT immunoreactivity and mutation status was assessed in 41 ALM and 25 MM patients, using polyclonal rabbit anti-human KIT/CD117 antibody, and PCR was performed for KIT gene exons 9,11,13,17 and 18. Of these, 19 ALM and 15 MM patients had matched primary and metastatic lesions. Phosphorylated extracellular regulated kinase (P-ERK) was investigated in a subset of 8 ALM and 8 MM matched primary/metastatic pairs by immunohistochemistry. Results: Heterogeneous KIT immunoreactivity was observed in both primary and metastatic lesions. Mutations were present in four of 41 ALM (10%) and five of 25 MM (20%) patients. Only vulvar mucosal samples carried KIT mutations in contrast to sinonasal lesions (p= 0.0109). In KIT-mutated tumors, the mutations were present in KIT expressing as well as KIT negative cells, as shown by Laser Capture Microdissection (LCM). P-ERK expression was preferentially found in metastases. Conclusions: KIT expression is heterogeneous and shows no relationship with disease progression and mutation status, therefore KIT immunoreactivity is not a valuable marker for treatment decisions. In mucosal melanoma patients, KIT mutation frequency is significantly associated with anatomic location. Vulvar melanoma patients are prone to carry activating KIT mutations and thus may be more likely to benefit from KIT-targeted therapies than other subtypes. Therefore, especially vulvar melanoma patients should be screened for activating KIT mutations.
MicroRNA alterations associated with BRAF status in melanoma.
Authors: Ma Michelle W. , Farhadian Joshua Andrew , Friedman Erica Brooke , Vega-Saenz de Miera Eleazar , Hanniford Douglas , Segura Miguel F. , Berman Russell S. , Shapiro Richard L. , Pavlick Anna C. , Zavadil Jiri , Hernando Eva , Osman Iman 
Published: March 2020

8565 Background: We hypothesize that BRAF mutations result in microRNA (miRNA) alterations which contribute to orchestrating the mutant BRAF’s oncogenic effects in melanoma. Our study is the first to examine the association between the BRAF mutation status in primary melanomas and the expression of miRNAs that target known tumor suppressors. Methods: 84 prospectively accrued melanoma patients at New York University Langone Medical Center were studied. DNA and total RNA were extracted from consecutive sections of formalin-fixed paraffin-embedded primary tissues. BRAF mutation status was determined by DNA sequencing. RNA was hybridized to miRCURY miRNA microarrays containing 1314 probes. Normalized miRNA data were analyzed using the t-test (p<0.05) to identify differentially expressed miRNAs between BRAFmut vs. BRAFwt cases. Those with an average fold change (FC) > 2 were selected for predicted (TargetScan, PicTar) and validated (miRWalk) gene target analysis, and overlapping genes targeted by ≥ 2 miRNAs were analyzed using pathway-mapping algorithms (KEGG, BioCarta, PANTHER). Results: 48 (57%) primaries were BRAFwt and 36 (43%) were BRAFmut (26 V600E, 4 V600K, 1 V600R, 1 V600D, 4 other). 30 miRNAs met the criteria for statistically significant differential expression and FC thresholding: let-7i, miR-23c, -26a/b, -27b, -34a, -98, -126*, -141, -148a, -181b, -195, -199a-3p, -199a/b-5p, -200a/b/c, -203, -205, -455-3p, -491-3p, -606, -641, -646, -1297, -4301; miRPlus-C1070, -C1110, -G1246-3p (average FC: 2.3-3.5, all increased in BRAFmut vs. BRAFwt). Predicted and validated target gene analysis revealed 317 genes, of which 110 (35%) were convergent targets of ≥ 2 miRNAs. Pathway analyses of the predicted, validated, and convergent target genes pointed to the potential impact of BRAFmut-associated miRNAs on known tumor suppressors FAS, PTEN, and TNF and the p53 pathway. Conclusions: Differentially expressed miRNAs in BRAFmut vs. BRAFwt primaries target genes with known roles in melanoma biology and/or treatment response. These miRNAs warrant further study as potential effectors of the BRAFmut oncogenic program.
Immunological and biological changes during ipilimumab (Ipi) treatment and their correlation with clinical response and survival.
Authors: Simeone Ester , Gentilcore Giusy , Romano Anna , Daponte Antonio , Caraco Corrado , Grimaldi Antonio M , Curvietto Marcello , Esposito Assunta , Queirolo Paola , Mozzillo Nicola , Ascierto Paolo Antonio 
Published: March 2020

8573 Background: Ipilimumab (Ipi) is approved in the US as first and second line therapy in patients with metastatic melanoma (MM) and in MM patients with previous therapy in the EU, based on an overall survival benefit shown in a phase III study (Hodi, NEJM 2010). To date, no clinical parameter has been found to be predictive for response to treatment and only few immunologic changes have been identified as potential candidates. Methods: From June 2010 to November 2011 we treated in the Expanded Access Program with Ipi at 3 mg/kg, 95 pre-treated metastatic melanoma patients. The median age was 58 yrs (range 17-84); 10 pts (10,5%) were stage IIIc inoperable, 2 pts (2,1%) stage M1a, 4 pts (4,2%) stage M1b, and 79 pts (83,2%) stage M1c. 30/95 pts had brain metastases and 1/95 spinal cord metastases. All 95 patients were evaluable for response (DCR = CR+PR+SD according the irRC), overall survival, safety, including changes in LDH, CRP (C-reactive protein) and lymphocyte populations (CD4+,CD4CD25+,FOXP3/T-Reg cells). PBMC and sera were collected at week 0, 4, 7, 10 and 12. Results: We found a statistical significant decrease of LDH, CRP and FOXP3/T-Reg cells (p<0.0001; χ2 and Mann-Whitney), and an increase of lymphocyte count (p<.0001) in the responders group. These differences were also correlated to survival (log-rank test). No differences were observed for CD4+ and CD4+CD25+ between responders and non-responders (p=0.39;p=0.83; Mann-Whitney). An ORR of 22.1% (1CR+20PR; 95% CI 13.8-30.4) and a DCR at week 24 of 37.9% (36/95; 28.1-47.6, 95% CI) were observed. Median overall survival was estimated of 7.8 months (95%CI:5.0-10.6), with a p value not evaluable at the moment of the analysis due to insufficient follow-up because of long-term survival. Adverse events were registered in 40% (38/95) of patients and the most frequent were of grade 1 and 2 (pruritus 57.9%; rash 5.3%; thyroditis 5.3%). Conclusions: The decrease of LDH, CRP and T-Reg cells during Ipi treatment suggest these parameters should be further explored as potential predictive markers for response and survival. Given the potential clinical utility of these findings, these data warrants further prospective validation in a randomized trial.
Metastatic basal cell carcinoma: Differences in survival by site of spread.
Authors: McCusker Margaret Elizabeth , Hou Jeannie , Wang Lisa , Yue Huibin , Hauschild Axel 
Published: March 2020

8585 Background: Basal cell carcinoma (BCC), the most common skin cancer, rarely metastasizes. Consequently, the epidemiology of metastatic BCC (mBCC) is poorly characterized. The largest published mBCC review includes 170 cases reported from 1894–1980 (von Domarus H, Stevens P. J Am Acad Dermatol 1984;10:1043-60). Targeted therapies with hedgehog signaling pathway inhibitors, such as vismodegib, are currently being developed to treat this rare disease. The objective of the current analysis was to provide an updated description of the epidemiology and survival outcomes for mBCC. Methods: A PubMed literature search was conducted for mBCC case reports published in English during 1981–2011. mBCC cases that met the following criteria were evaluated: primary BCC located on skin; primary tumor and metastasis confirmed by pathology, and metastasis was not the result of direct tumor spread. Only cases with survival data giving dates (month and year) or durations were included in the analysis. Differences between groups were assessed with t tests and χ2 tests as appropriate. Survival was assessed with Kaplan–Meier (K-M) methods. Results: We identified 101 mBCC cases that met the selection criteria; 38 patients (38%) had lymph node-only disease (LD) and 63 (62%) had distant metastases (DM). In both groups, males predominated. DM patients were younger at mBCC diagnosis (mean 59 vs 66 y for LD). Among 96 cases with treatment data for metastatic disease, more DM patients received chemotherapy (25% vs 9% for LD), but more LD patients underwent surgery (85% vs 52% for DM). Survival duration ranged from 0 to 120+ months in all patients. The overall 1-y probability of survival after mBCC diagnosis was 75.7% (95% CI 67.2–84.2%), with lower survival in DM patients (69.1%; 95% CI 57.5–80.7%) vs LD patients (86.5%; 95% CI 75.5–97.5%). Conclusions: Patients with LD and DM mBCC may have different clinical courses and outcomes. Based on published case reports, DM patients were younger at mBCC diagnosis and had a lower 1-y survival probability vs LD patients. To our knowledge, these are the first K-M survival estimates reported for an mBCC case series of this size. These data help to provide a historical context for novel mBCC therapies under development, such as vismodegib.
Clinical experience with atypical spitzoid tumors in patients younger than age 18: Does fluorescence in situ hybridization predict lymph node metastasis?
Authors: Stanelle Eric J. , Busam Klaus J. , Rich Barrie S. , Coit Daniel G. , La Quaglia Michael P. 
Published: March 2020

8586 Background: Determining the malignant potential of atypical spitzoid melanocytic proliferations can be diagnostically challenging, and many patients are therefore managed as if they had melanoma. However, few studies focus specifically on atypical spitzoid tumors (AST). Further, cytogenetic analysis using fluorescence in situ hybridization (FISH) has been used to determine malignant potential. We reviewed our institutional experience to determine staging and clinical outcomes, and the correlation between FISH findings and regional nodal positivity in patients with AST. Methods: With IRB approval, we retrospectively reviewed all patients aged <18 years treated for AST from 1981-2011 to determine staging variables, outcome, and the results of 4-probe FISH assay. A total of 44 patients with a median age of 11.5 years were identified. All pathology was re-reviewed by a single dermatopathologist. Staging was based on 2010 AJCC criteria. Correlations were determined using either the Pearson or Spearman correlation coefficient. Results: Median lesion thickness was 2.8 mm (range: 0.55-12) and eight (18%) lesions met spitzoid melanoma criteria. Median followup was 4.1 years for all patients (5.5 years for spitzoid melanomas). Thirty-nine patients (89%) underwent sentinel lymph node biopsies (SLNB) and 15 (38%) were positive. Lymph node (LN) positivity correlated with tumor thickness (p=0.002), stage (p=0.001), and mitotic rate (p=0.05). FISH was available for 17/39 patients who had SLNB; sensitivity and specificity for LN metastases were 50% and 54%, respectively. Of 15 patients with a positive SLNB, 12 underwent completion LN dissection, three of which were positive for 1, 2, and 3 additional nodes, respectively. There were no recurrences or disease-related deaths. Conclusions: Traditional indicators of thickness and stage predicted nodal involvement in pediatric patients with AST. However, FISH results did not predict nodal status and should not be used to guide management. With 100% disease specific survival and no recurrences, AST should be considered a separate entity from melanoma and complete excision may be sufficient therapy.
A phase II, multicenter, open-label study of YM155 plus docetaxel in subjects with stage III (unresectable) or stage IV melanoma.
Authors: Steinberg Joyce Leta , Bedikian Agop Y. , Ernst D. Scott , Chmielowski Bartosz , Redman Bruce , Keating Anne Therese , Jie Fei , Chen Caroline , Weber Jeffrey S. , Kudchadkar Ragini Reiney ,   YM155-CL-034 Study Group
Published: March 2020

8587 Background: Survivin is a microtubule-associated protein implicated in both preservation of cell viability and regulation of mitosis in tumor cells. It is over-expressed in melanoma, breast, and non-Hodgkin’s lymphoma. YM155 is a first in class survivin inhibitor. Methods: The study had 2 parts: Part 1 established the dose of docetaxel that was tolerable in combination withYM155 at 5 mg/m2/day continuous infusion over 168 hours q 3 weeks. Part 2 utilized the dose of docetaxel established in Part 1 to further evaluate the tolerability and activity of the combination. The primary endpoint was 6-month progression-free survival (PFS). Secondary endpoints were overall response rate, 1-year overall survival (OS), time from first response to progression, clinical benefit rate, time to response, and safety. Results: 64 patients with metastatic melanoma were treated with docetaxel followed by continuous infusion YM155. 7 patients were treated with 100mg/m2 of docetaxel and 57 patients were treated with 75mg/m2 of docetaxel. Median age was 59, with 44 men and 20 women treated. 6-month PFS per Independent Review Committee (IRC) was 34.8% (95% CI 21.3 – 48.6%). Overall objective response rate per IRC was 12.5%, with no complete responses (CR) and 8 patients with partial responses (PR). The Stable disease (SD) rate was 51.6%, leading to a clinical benefit rate (CR + PR + SD) of 64.1%. Estimated 1-year overall survival is 50.5%. 87.5% of patients experienced a Grade 3 (G3) or Grade 4 (G4) event attributable to either YM155 or docetaxel. The clinically pertinent G3 or 4 toxicities occurring in greater than 5% of patients treated included neutropenia (59.4%), febrile neutropenia (12.5%), mucositis (9.4%), fatigue (7.8%), diarrhea (6.3%), and dehydration (6.3%). There were 3 deaths on study, all attributable to disease progression. Conclusions: YM155 is a novel agent that shows modest activity when combined with docetaxel in patients with melanoma. YM155 was generally well tolerated, but the pre-determined primary endpoint for efficacy was not achieved.
Pharmacodynamic activity of selumetinib to predict radiographic response in advanced uveal melanoma.
Authors: Carvajal Richard D. , Ambrosini Grazia , Wolchok Jedd D. , Chapman Paul B. , Dickson Mark Andrew , D'Angelo Sandra P. , Bluth Mark J. , Paucar Daniel , Fusco Anne , Bohr David , Roman Ruth Ann , Montefusco Mary , Doyle L. Austin , Marr Brian , Abramson David H. , Chou Joanne F. , Panageas Katherine , Schwartz Gary K. 
Published: March 2020

8598 Background: Functionally activating mutations (mut) in Gnaq or Gna11, genes that encode for widely expressed G-protein alpha subunits, are early oncogenic events in uveal melanoma (UM) development and result in activation of the MAPK pathway. We previously demonstrated effective pathway inhibition with selumetinib (AZD6244, ARRY-142866) in UM cell lines, with decreased viability associated with pERK and cyclinD1 suppression (Ambrosini, AACR 2010). Methods: Using paired metastatic tumor biopsies from patients (pts) with radiographically progressing UM treated with selumetinib 75 mg BID on a phase II trial (NCT01143402), we correlated MAPK pathway inhibition with radiographic tumor regression and clinical benefit. Biopsies were performed at baseline and after 14 +/-1 days of treatment. Western blotting was performed for pERK and cyclinD1, and quantitated by densitometry. Response (RECIST 1.1) was assessed at baseline, week (wk) 4, wk 8, and q8wks subsequently. Radiographic regression was defined as greatest percentage shrinkage from baseline. Clinical benefit was defined as RECIST response or stable disease ≥16wks. Results: Paired tumor biopsies were assayed from 18 pts: median age 60 (range 47-81), M:F 11:7, median 1 prior therapy (range 0-2), 17 with liver involvement, Gnaq mut:Gna11 mut:wild-type 8:9:1. Radiographic regression was observed in 5 pts, with 2 achieving partial responses. 4 pts were on study ≥16wks (16+, 20, 25, 31 wks), with one currently on study at 11+ wks. Median pERK and cyclinD1 as measured by densitometry decreased by 48% (p=.03) and 76% (p=.03), respectively. Radiographic regression correlated with suppression of pERK (Spearmen’s rank correlation; p=0.04) but not cyclinD1 (p=0.38). A trend towards pERK suppression correlating with clinical benefit was observed (p=.07) with each of the 5 pts achieving PR or SD ≥16wks having a decrease of ≥30% in pERK from baseline. Conclusions: Selumetinib can inhibit pERK and cyclinD1 in UM and can result in tumor shrinkage. Sustained inhibition of pERK inhibition at day 14 may be predictive of benefit. Further evaluation of MEK inhibition in this disease is warranted.
Exhibition of a stem-cell like phenotype with the expression of CD271 in drug-resistant melanoma cells.
Authors: Lieser Elizabeth Ann Teresa , Leontovich Alexey A , Holtan Shernan G. , Nevala Wendy Kay , Markovic Svetomir 
Published: March 2020

8599 Background: Chemotherapy resistance is a common, difficult problem for melanoma patients needing long term care and those suffering from recurrence. Neural growth factor receptor (NGFR), also known as CD271, is commonly expressed on mesenchymal stem cells and is important for development, survival and differentiation of cells. This has previously shown to be expressed on melanoma stem cells. We hypothesized that treatment with carboplatin enriches the population of tumor cells for cancer stem cells (CSC) as a mechanism of chemotherapy resistance. Methods: Core tumor samples from 118 patients with metastatic melanoma were obtained from formalin-fixed paraffin embedded specimens. Slides were stained using an anti-CD271 antibody for IHC. A375 melanoma cells were treated with increasing concentrations of carboplatin for approximately 1 year. WT, 6 mg/mL and 10 mg/mL resistant cells were tagged with CD271-PE antibody and run on a flow cytometer. These cells were then tested for gene expression by microarray using an Affymetrix human 133 plus 2.0 chip, and analyzed on Partek software. Pathway analysis of these carboplatin resistant cells was preformed using Ingenuity. Results: Virtually all melanoma tumor samples stained positively for CD271 by IHC, with a median of 50% cells within tumor samples expressing the cytoplasmic marker. Flow cytometry demonstrated an increase in the percentage of cells expressing CD271 as resistance to carboplatin increased. Wild type A375 cells contained 30% positive CD271, A375 6mg/mL resistant contained 42% positive and A375 10mg/mL resistant showed 66% positive. Microarray also exhibited a direct correlation between CD271 gene expression with increasing carboplatin resistance. Functional ontology enrichment revealed development and regulation of the EMT pathway as an important process impacted by our resistant cell line. Conclusions: The data suggests that as melanoma cells become resistant to certain chemotherapy drugs they essentially loose their differentiated phenotype and become more stem cell-like. Monitoring melanoma expression of CD271 could help individualize therapy and anticipate chemotherapeutic resistance in this high-risk group of patients.