bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effect of the specific P53 stabilizer CBS9106 on multiple myeloma (MM)
Authors: Ikeda H. , Hideshima T. , Perrone G. , Okawa Y. , Raje N. , Kawabe T. , Sakakibara K. , Saito N. , Richardson P. , Anderson K. 
Published: August 2020

8601 Background: The mutations of P53 tumor suppressor protein are associated with progressive in Multiple Myeloma (MM), conversely, stabilization of P53 leads to cell cycle arrest and apoptosis. In this study, we examined p53 protein expression and demonstrated the effect of P53 stabilization using a novel specific P53 stabilizer CBS9106 in MM. Method: We examined P53 protein expression using Immunoblot analysis, as well as the growth inhibitory effect of CBS9106 in MM cell lines and primary tumor cells from MM patients. We also defined whether CBS9106 can overcome the growth promoting effect of exogenous cytokines and bone marrow stroma cells (BMSCs) using [3H]-thymidine uptake assay. Results: Expression of P53 protein was observed in 3/3 primary tumor cells from MM patients and 6/6 MM cell lines. CBS9106 at low nM levels triggered cytotoxicity against p53 wild type MM cell lines and primary tumor cells from MM patients, associated with phosphorylation of P53 (serine15 and 20). In contrast, CBS9106 did not affect the survival of normal peripheral blood mononuclear cells from healthy volunteers at concentrations as high as 10 μM. This agent also induced G1 cell cycle arrest, followed by apoptosis associated with cleavage of caspase-3, -8, -9 and PARP. Neither growth stimulating cytokines (IL-6 and IGF-1) nor BMSCs protected against apoptotic effect of CBS9106. Moreover, we demonstrate that combination of CBS9106 with MDM2 inhibitor Nutrin3 or proteasome inhibitor bortezomib induces synergistic anti-MM activity in both P53 wild type MM cell lines and primary tumor cells from MM patients. Conclusions: Stabilizing P53 by CBS9106 represents a novel promising p53-based therapy in MM. These results provide the preclinical framework supporting evaluation of CBS9106 in clinical trials to improve patient outcome in MM. No significant financial relationships to disclose.
Impact of dexamethasone responsiveness on long-term outcome in patients with newly diagnosed myeloma
Authors: Sinha S. , Rajkumar S. V. , Lacy M. , Hayman S. , Baudi F. , Dispenzieri A. , Lust J. , Greipp P. , Gertz M. , Kumar S. 
Published: August 2020

8603 Background: Steroids, primarily dexamethasone (Dex), remains an integral part of the therapy for myeloma. Prior to the advent of novel agents, Dex as a single agent was demonstrated to produce overall response rates of around 50%. The toxicity associated with pulsed high dose Dex has led to this approach being abandoned. This study investigates whether responsiveness to Dex correlates with biological qualities of the plasma cell clone that predicts outcome. Methods: We identified from our prospectively maintained database, patients with newly diagnosed myeloma, who were seen at Mayo Clinic within 30 days of diagnosis and were treated with single agent Dex. The M-protein measurements and responses were assessed using the IMWG criteria. Results: 182 patients seen between March 1998 and June 2007, initially treated with single agent Dex for at least 4 weeks, were included in the analysis. The median age at diagnosis was 63 (range; 39–81) and 59% were male. The actuarial median survival for the entire group was 55 months. The median duration of Dex therapy was 15 weeks (range; 4–193). Ninety one (50%) had a partial response or better and 11 (6%) were not evaluable, with the remaining patients (44%) having less than PR or progression. 10 patients (5%) achieved a VGPR or CR. The median overall survival from diagnosis for the 91 responders was 75 months compared to 71 months for the remaining patients; p = 0.6. There was no correlation between baseline disease characteristics and Dex responsiveness. One hundred thirty (70%) patients proceeded to an autologous SCT after a median of 15 weeks (range 1–46 months) of Dex. While the median overall survival was longer among the group of patients who underwent SCT, among either group, there were no survival benefit between those with a PR or better to Dex compared to the rest. Conclusions: Among this group of patients with newly diagnosed myeloma treated with single agent Dex, failure to respond to Dex induction therapy did not impact overall survival. Failure to respond to single agent steroid does not have an adverse impact on outcome in newly diagnosed myeloma. [Table: see text]
Prognostic impact of cancer-testis antigen expression in primary cutaneous melanoma
Authors: Cebon J. S. , Svobodova S. , Browning J. , Davis I. D. , Scolyer R. , Murali R. , Thompson J. F. , Deb S. , Azad A. , MacGregor D. 
Published: August 2020

9004 Background: Cancer-testis antigens (CTAg) are epigenetically regulated molecules expressed in many cancers including melanoma. Although functional studies are limited, they are often immunogenic making them attractive targets for immunotherapy. In normal tissues expression is restricted to germ cells and a small range of other tissues such as trophoblast. Previous studies have shown that CTAg expression increases with disease progression. We investigated whether the expression of three CTAgs, against which vaccines have been developed, may have prognostic significance in early stage melanoma. Methods: 233 AJCC Stage II melanomas were analyzed for expression of MAGE-A1, MAGE-A4 and NY-ESO-1 by immunohistochemistry. The relationship between CTAg expression, clinico-pathological features and relapse free survival (RFS) from initial diagnosis were correlated. Mutivariate analysis using known prognostic factors and CT Ag expression in the model were used to calculate hazard ratios. Results: All three CTAg were significantly co-expressed with each other (P=0.0001). RFS was reduced if tumors expressed any of these CTAgs (CTAg+ve). Median RFS for patients with CTAg+ve tumors was 45m versus 72m for those with CTAg-ve tumors (P=0.008, logrank test). Univariate analysis demonstrated that the impact of CTAg expression on RFS was comparable in magnitude to ulceration, Breslow thickness and mitotic rate, currently accepted prognostic factors. Multivariate analysis demonstrated CTAg expression, ulceration and thickness but not mitotic rate were independently associated with poorer RFS ( Table ). Conclusions: CTAg expression in cutaneous primary melanoma has impact on prognosis comparable to Breslow thickness ulceration and mitotic rate. Further study into their function and the impact of clinical targeting is warranted. [Table: see text] No significant financial relationships to disclose.
Long-term follow-up of patients with minimal sentinel node tumor burden (< 0.1mm) according to Rotterdam criteria: A study of the EORTC Melanoma Group
Authors: van Akkooi A. C. , Rutkowski P. , van der Ploeg I. M. , Voit C. A. , Hoekstra H. J. , Nieweg O. E. , Schäfer-Hesterberg G. , Nowecki Z. I. , de Wilt J. H. , Eggermont A. M. 
Published: August 2020

9005 Background: Many studies have identified Sentinel Node (SN) tumor burden as a prognostic factor for additional non- SN (NSN) positivity and / or disease-free (DFS) and melanoma specific survival (MSS). It remains unclear if pts with minimal SN tumor burden can safely be managed without Completion Lymph Node Dissection (CLND). Pts with minimal SN tumor burden might be at risk for late recurrences (> 5 years). Methods: Slides of 595 SN positive patients were reviewed for this pan-European study collaboration in 5 major centers. Slides were reviewed for the microanatomic location and SN tumor burden according to the Rotterdam Criteria (< 0.1mm, 0.1 - 1.0mm and > 1.0 mm) for the maximum diameter of the largest metastasis. MSS, DFS and distant metastasis-free survival (DMFS) rates were calculated, as was NSN positivity. Results: In 595 SN positive pts, the mean and median Breslow thickness was 4.73 and 3.5 mm. Ulceration was present in 51% of melanomas. 67 pts had metastases < 0.1 mm (11%), 226 pts (38%) had 0.1 - 1.0 mm metastases and 302 pts had metastases > 1.0 mm (51%). Mean and median follow-up was 48 and 40 months for all patients (range 1 - 172). Patients with metastases < 0.1 mm had mean and median follow-up of 61 and 57 months, 46% (31pts) had follow up > 5 years and 25% (17 pts) had follow-up longer than 80 months (range 3 - 132). 5-year MSS rates were 94% for metastases < 0.1 mm, 70% for 0.1 - 1.0 mm and 57% for > 1.0 mm (p<0.001). 5-year DMFS rate was 91% for metastases < 0.1 mm. NSN positivity occurred in 5% of < 0.1mm, 17% of 0.1 - 1.0 and 29% of metastases > 1.0 mm (p<0.001). Conclusions: This large multicenter experience (n=595) has demonstrated that long-term follow-up of melanoma patients with minimal SN tumor burden (< 0.1 mm) indicates very low relapse rates and excellent MSS, seemingly identical to SN negative patients. With prolonged follow-up, an increase in the occurrence of relapses of any kind between 5 and 10 years follow up has not been identified, and excellent 10-year survival rates are expected. No significant financial relationships to disclose.
Ulceration of primary melanoma and responsiveness to adjuvant interferon therapy: Analysis of the adjuvant trials EORTC18952 and EORTC18991 in 2,644 patients
Authors: Eggermont A. M. , Suciu S. , Testori A. , Patel P. , Spatz A. 
Published: August 2020

9007 Background: Ulcerated (Ulc) melanomas have a worse prognosis than non-ulcerated (N-Ulc) melanomas. Ulc and N-Ulc primaries have different stromal characteristics and gene profiles reflecting differences in biology. We analyzed outcome after adjuvant interferon (IFN) therapy in the 2 largest phase III trials (EORTC18952 and 18991) ever conducted in stage IIB-III melanoma patients (pts). Methods: EORTC18952 compared IFNα-2b (10 MIU, sc, qd, 5 days/wk, for 4 wks) followed by either 10MIU,sc, tiw for 12 mts, or 5MIU for 24 mts) with observation in 1,388 stage IIB-III pts (Lancet 2005;366). EORTC18991 evaluated pegylated IFNα-2b (6 μg/Kg, 1×/wk, for 8 wks followed by 3μg/Kg, 1×/wk for up to 5 yrs) versus observation in 1,256 stage III pts (Lancet 2008;372). Using meta-analytical methods, predictive value for Ulc on the value of IFN on relapse-free survival (RFS), distant metastasis-free survival (DMFS), and overall survival (OS) was assessed, overall, and according to stage (IIB, III-N1 or N2=microscopic or macroscopic-nodal disease). Results: Overall, the comparison (PEG-)IFNα-2b versus observation regarding RFS, DMFS, and OS led to a reduction in the hazard ratio (HR) of -16% (SE=5%), -13%(5%), and -8%(5%). Among 2,644 pts randomized, 849 had Ulc primaries, 1,336 N-Ulc primaries, and 459 Ulc unknown. In Ulc group the impact was much greater than in N-Ulc group for RFS (Test For Interaction: p=0.02), DMFS (p<0.001), and OS (p<0.001). The greatest reductions occurred in pts with Ulc and stages IIB/III-N1 In N-Ulc pts reduction was absent. Consistency in the treatment impact was seen in both trials. Conclusions: The post hoc analyses of EORTC1892 and EORTC18991 indicate strongly that pts with an Ulc primary are far more sensitive to IFN than pts with N-Ulc primaries. This hypothesis will now be tested in the EORTC18081 trial, which compares PEG-IFNα-2b versus observation in pts with Ulc primaries ≥ 1mm. [Table: see text] [Table: see text]
Correlation of ultrasound criteria for detection of melanoma metastases in the sentinel lymph node (SN) with tumor burden and survival
Authors: Voit C. A. , van Akkooi A. C. , Schaefer-Hesterberg G. , Schoengen A. , Sterry W. , Eggermont A. M. 
Published: August 2020

9015 Background: We demonstrated that US guided FNAC (fine needle aspiration cytology) prior to SN biopsy can identify up to 65% of SN-positive patients (EJC, 2007; 5(6):11: abstr 3BA). We presented US patterns of SN-involvement at ASCO 2008 (JCO 26: 2008(suppl; abstr 9014). Here we show how these patterns correlate with tumor burden, progression of disease and survival. Methods: Prior to SN-biopsy patients (pts) underwent lymphoscintigraphy followed by US-exam. US images were prospectively scored for 6 morphologic criteria. FNAC was performed in suspicious US. All pts underwent a SN biopsy. Final SN pathology was the gold standard. Sensitivity, specificity and negative/positive predictive value (NPV and PPV) of combinations of US patterns were calculated and correlated with tumor burden and survival. Hazard ratios (HR) were calculated for the patterns by multivariate analysis. Results: Since 2001 850 consecutive pts have been included into a prospective database. Median Breslow thickness of the first 400 stage I/II melanoma pts was 1.8 mm, median follow-up 42 months. Balloon Shape (BS) & Loss of Central Echoes (LCE) are often linked (up to 83%) and are late signs correlating with high tumor load. In contrast the presence of Peripheral Perfusion (PP) is an early sign, correlating with small tumor load. PP and/or BS and/or LCE together raise the sensitivity of US alone to > 80%, spec.80%, PPV 52%, NPV of 94% (p<0.001). Overall Survival of neither vs. Peripheral Perfusion (PP) only vs. BS/LCE (with or without PP) was 93% vs. 87 % vs. 56 % and Distant Metastasis-Free Survival was 74% vs. 60% vs. 26%. BS/LCE was a late sign correlating with high tumor load, fast progression and a high HR (5.50). PP alone was an early sign correlating with small tumor load, slow progression, and a low HR (2.19). Conclusions: BS and/or LCE indicate high tumor load, PP alone indicates small tumor load in the SN. These US-criteria identify the SN tumor burden correctly prior to SN biopsy in 75% - 90% of pts. They are independent prognostic factors for survival. No significant financial relationships to disclose.
Serum vitamin D levels, VDR, and survival from melanoma
Authors: Newton Bishop J. A. , Beswick S. , Randerson-Moor J. , Chang Y. , Affleck P. , Elliott F. , Elder D. , Barrett J. , Bishop T. 
Published: August 2020

9016 Background: Vitamin D has pleiotropic effects, which may moderate the interaction between patients and their tumors. Two studies were carried out to test the hypothesis that higher vitamin D levels reduce the risk of relapse from melanoma. Methods: A pilot retrospective case-control study in 271 melanoma patients suggested that vitamin D may protect against recurrence of melanoma. We then tested these findings in a survival analysis in a cohort of 872 cases recruited to the Leeds Melanoma Cohort (median follow up of 4.7 years). Results: Pilot study Results suggested that taking vitamin D reduced the risk of relapse from melanoma (OR 0.6, 95% CI: 0.4, 1.1). Non-relapsers had higher mean 25-dihydroxyvitamin D3 levels than relapsers (49 nmol/L compared with 46, p=0.3). Cohort study Higher 25-dihydroxyvitamin D3 levels were associated with lower Breslow thickness at diagnosis and were independently protective of relapse and death: hazard ratio (HR) for relapse free survival (RFS) 0.76, 95% CI: (0.64, 0.96), for a 20nmol/L increase in serum level. There was evidence of interaction between the vitamin D receptor (VDR) BsmI genotype and serum 25-dihydroxyvitamin D3 levels on RFS. Conclusions: The pilot study provided preliminary evidence for a role for vitamin D in outcome from melanoma. The cohort study provided further evidence that higher 25-dihydroxyvitamin D3 levels, at diagnosis, were associated both with thinner tumors and better survival, independent of Breslow thickness, from melanoma. Melanoma patients should avoid vitamin D deficiency. Further studies are needed to establish optimal serum levels for melanoma patients. No significant financial relationships to disclose.
Selective activity of dasatinib for the most common KIT mutation in melanoma (L576P)
Authors: Woodman S. E. , Trent J. C. , Stemke-Hale K. , Lazar A. , Pricl S. , Pavan G. M. , Papadopoulos N. , Hwu P. , Mills G. B. , Davies M. A. 
Published: August 2020

9019 Background: Point mutations in the KIT receptor tyrosine kinase gene have recently been identified in melanomas from mucosal, acral lentiginous (AL), and chronically sun-damaged (CSD) sites. An improved understanding of the molecular characteristics of melanoma-prevalent KIT mutations may lead to more effective therapeutic approaches. Methods: Human melanoma cell lines were screened by mass-spectroscopy based genotyping for KIT mutations, and a cell line with an endogenous L576P KIT mutation was identified. The cell line was treated in vitro with a panel of small molecule KIT inhibitors and the effects on cell viability were quantified. Molecular modeling of the interaction of the inhibitors with the KIT L576P mutant protein was determined to estimate binding affinity. PET/CT studies were performed on patients with mucosal melanoma harboring the L576P mutation pre- and post-dasatinib treatment. Results: We have identified the first human melanoma cell line with an endogenous L576P mutation, the most common KIT mutation in melanoma. In vitro testing demonstrated that this cell line is resistant to imatinib, nilotinib and sorafenib (0 - 1 uM), KIT inhibitors shown to be effective in non-melanoma cells with other KIT mutations. However, the mutant cell line was inhibited by dasatinib at concentrations as low as 10 nM, and was significantly more sensitive than melanoma cell lines with wild-type KIT (p = 0.02). No difference in sensitivity to Src inhibitors was observed, supporting that this sensitivity was due to KIT inhibition. Molecular modeling demonstrated that the L576P mutation induces structural changes in KIT that reduce the affinity for imatinib but not for dasatinib. Two metastatic melanoma patients with the L576P KIT mutation were treated with dasatinib, including one patient previously treated with imatinib. Both patients had marked reduction and elimination of tumor FDG- avidity by PET imaging after dasatinib treatment. Conclusions: This data supports that dasatinib has a selective inhibitory effect against the most common KITmutation in melanoma and has implications for the development of AL, CSD, and mucosal melanoma treatment. [Table: see text]
Quantitative assessment of AKT activation in melanoma
Authors: Davies M. A. , Stemke-Hale K. , Calderone T. , Deng W. , Lazar A. , Prieto V. G. , Aldape K. , Mills G. B. , Gershenwald J. E. 
Published: August 2020

9022 Background: Activating mutations of BRAF are highly prevalent in melanoma. However, multiple lines of evidence suggest that other pathways must also contribute to this disease. Activation of the PI3K-AKT pathway has been implicated in melanoma by mutations of NRAS and PTEN. Little direct information is known about the activation of the PI3K-AKT signaling pathway in melanoma, particularly in metastases. Methods: Proteins isolated from 99 frozen melanoma tumors were measured by reverse phase protein arrays (RRPA). A total of 53 proteins were assessed using validated antibodies. Activating mutations were assessed by mass spectroscopy-based genotyping. Activation of AKT (phospho-AKT) was compared to mutations and anatomic sites. Results: Samples from 75 regional metastases (LN or in-transit) and 24 distant metastases were analyzed. Technical replicates (same lysate) and biological replicates (same tumor, different lysates) demonstrated average Pearson correlation coefficients (r) of > 0.90, supporting the high technical quality of the analysis. A positive correlation was observed between levels of p-AKT and known AKT substrates p-GSK3 and p-TSC2 (p< 0.001 for both), and a negative correlation with PTEN (p < 0.01), supporting the maintenance of phosphorylation events during sample processing. Relative differences in p-AKT by RPPA were also confirmed by immunohistochemistry of representative tumors. Tumors with BRAF mutations had higher levels of p-AKT than tumors with NRAS mutations (p = 0.03). Indeed, tumors with NRAS mutations had p-AKT levels similar to tumors wild-type for BRAF and NRAS (p =0.73). Detailed analysis demonstrated that all tumors with elevated p-AKT had low PTEN expression. Similar results were seen in human melanoma cell lines. Analysis of distant metastases demonstrated that brain metastases had higher levels of p-AKT, p-GSK3, and p-TSC2, and lower levels of PTEN, compared to metastases to the lung or liver. Conclusions: AKT activation in melanoma correlates with PTEN expression, and does not correlate with NRAS mutation. Activation of the PI3K-AKT pathway may contribute to the aggressiveness of brain metastases. These findings have clinical implications for targeting this pathway, and demonstrate the feasibility and potential of RPPA analysis of signaling pathways in melanoma. No significant financial relationships to disclose.
Therapeutic vaccination with an autologous TriMix-Dendritic cell vaccine combined with sequential interferon alfa-2b in patients with advanced melanoma
Authors: Wilgenhof S. , Van Nuffel A. M. , Benteyn D. , Pierret L. , Heirman C. , De Coninck A. , Van Riet I. , Bonehill A. , Thielemans K. , Neyns B. 
Published: August 2020

9024 Background: Electroporation of dendritic cells (DC) with mRNA encoding fusion-proteins of a HLA-class II targeting signal and a melanoma associated antigen (MAA) together with mRNA encoding CD40 ligand, a constitutively active TLR4 and CD70 (TriMix) improves the immunostimulatory capacity of autologous DC. Methods: Following leukapheresis, immature DCs (derived from adherent PBMC cultured for 6 days in IL-4 / GM-CSF supplemented medium) are electroporated with mRNA encoding MAGE-A3, MAGE-C2, Tyrosinase and gp100 linked to DC-LAMP, and TriMix mRNA. TriMix-DC (12.5 106/antigen) are cryopreserved and administered by 4 ID-injections q2w, and q8w thereafter. After the 4th vaccination, interferon alfa-2b (IFN- a2b, 5 MIU TIW) is initiated. Immune monitoring is performed by skin biopsy of a vaccine injection site. Biopsies are investigated by IHC and by analyzing the activation (CD137+), cytolytic capacity (CD107a+), and cytokine release (IFN-γ and TNF-α) of DTH infiltrating T-cells in response to autologous EBV-B cells expressing MAA. Results: 29 pts (17M/12F; med age 49, range 28–75) with stage III/IV melanoma, nl LDH, and no CNS metastases were recruited. Vaccine related AE's (first 24 pts): gr2 local injection site reactions (all pts); fever & lethargy (gr2, 1 pt). Pts (20) who initiated IFN-a2b experienced constitutional side effects (gr3, 1 pt). Vaccine-specific DTH infiltrating T cells were documented post-vaccination in 13/17 pts (10/13 pts had a CD137+CD8+ and 2/13 pts a CD4+ response). Out of the 11 pts without evaluable disease, 2 had a local recurrence (salvaged by surgery). After a mFU of 7.8 mths (range 4.3–13.7) all pts remain disease-free. Out of the 13 pts with measurable disease, BOR (RECIST) was 8 SD and 5 PD; 1 pt with initial PD subsequently obtained a PR. Regression of metastases occurred in lung- (2 pts), orbita- (1 pt) and lymph node metastases (3 pts). After a mFU of 7 mths (range 1–14), the mPFS is 3,1 mths (95% CI 2,29–4,08); 4 pts remain progression-free after respectively 5, 8, 10 and 11 mths of follow-up. Conclusions: Therapeutic vaccination with TriMix-DC combined with sequential IFN-a2b is feasible, safe, immunogenic and associated with anti-tumor activity in patients with advanced melanoma. [Table: see text]