Citations

KEYNOTE-030 (ClinicalTrials.gov ID, NCT02083484) was a global expanded access program that allowed access to pembrolizumab, an antiprogrammed death 1 antibody, for patients with advanced melanoma before its regulatory approval. Patients with unresectable stage III/IV melanoma that progressed after standard-of-care therapy, including ipilimumab and, if BRAF V600 mutant, a BRAF inhibitor, were eligible to receive pembrolizumab 2 mg/kg every 3 weeks. Response was assessed by immune-related response criteria by investigator review. Adverse events (AEs) were graded according to the National Cancer Institute Common Terminology Criteria for Adverse Events, version 4.0. In the United States, 979 patients enrolled between April and September 2014. Of the 947 evaluable patients, 621 (65.6%) remained on treatment and transitioned to receive commercial pembrolizumab following approval by the Food and Drug Administration, whereas 326 (34.4%) discontinued, most commonly for disease progression (39.6%) or death (26.4%). Objective response rate was 14.5% (95% confidence interval, 12.2%–16.8%) in the treated population (n=947) and 22.1% (95% confidence interval, 18.8%–25.5%) in patients who had ≥1 response assessment reported (n=619). Twelve patients achieved complete response. One hundred eighty-one (19.1%) patients experienced ≥1 treatment-related AE, most commonly general disorders (8.0%), skin/subcutaneous tissue disorders (7.3%), and gastrointestinal disorders (6.4%); 29 (3.1%) patients experienced ≥1 grade 3/4 treatment-related AE. Immune-mediated AEs were also reported. There were no treatment-related deaths. The safety and efficacy observed in this expanded access program were consistent with those previously reported for similar populations and support the use of pembrolizumab for patients with advanced melanoma.

ABSTRACTChronic low back pain is a major cause of disability and health care costs. Effective treatments are inadequate for many patients. Animal models are essential to further understanding of the pain mechanism and testing potential therapies. Currently, a number of preclinical models have been developed attempting to mimic aspects of clinical conditions that contribute to low back pain (LBP). This review focused on describing these animal models and the main behavioral tests for assessing pain in each model. Animal models of LBP can be divided into the following five categories: Discogenic LBP, radicular back pain, facet joint osteoarthritis back pain, muscle‐induced LBP, and spontaneous occurring LBP models. These models are important not only for enhancing our knowledge of how LBP is generated, but also for the development of novel therapeutic regimens to treat LBP in patients. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:1305–1312, 2018.

Abstract Glioma stem cells (GSC) and epithelial–mesenchymal transition (EMT) are strongly associated with therapy resistance and tumor recurrence, but the underlying mechanisms are incompletely understood. Here, we show that S100A4 is a novel biomarker of GSCs. S100A4+ cells in gliomas are enriched with cancer cells that have tumor-initiating and sphere-forming abilities, with the majority located in perivascular niches where GSCs are found. Selective ablation of S100A4-expressing cells was sufficient to block tumor growth in vitro and in vivo. We also identified S100A4 as a critical regulator of GSC self-renewal in mouse and patient-derived glioma tumorspheres. In contrast with previous reports of S100A4 as a reporter of EMT, we discovered that S100A4 is an upstream regulator of the master EMT regulators SNAIL2 and ZEB along with other mesenchymal transition regulators in glioblastoma. Overall, our results establish S100A4 as a central node in a molecular network that controls stemness and EMT in glioblastoma, suggesting S100A4 as a candidate therapeutic target. Cancer Res; 77(19); 5360–73. ©2017 AACR.

Abstract Glioblastoma is the most common primary tumor of the CNS in adults, representing approximately 50% of all gliomas and 15% of primary brain tumors. The current standard of care for GBM is safe maximal surgical resection followed by radiotherapy and concurrent Temozolomide (TMZ), but median survival continues to be 15–16 months. Temozolomide is a second-generation DNA alkylating agent that induces thymine mispairing during DNA replication resulting in tumor cell G2/M phase arrest and autophagy, a standard care of therapy against GBM. While the success of TMZ in clinical trials showed great promise for its overall efficacy, emerging TMZ resistance make us to think more for the combination therapy. CD47, a tumor cell surface marker, plays as “don't eat me signal” through binding its receptor SIRPα on macrophages and the antibody against CD47, which blocks interactions of CD47 with SIRPα, has been shown to lead to tumor destruction. Furthermore, CD47 is a prognostic marker as its expression predisposes cancer patients to a poorer survival outcome. This has significant clinical implications since approximately more than 80% of patients with the most GBMs, overexpress CD47. Overexpression of CD47 is also associated with a decreased probability of survival in clinical cohorts of GBM. Our work and that of others demonstrate that CD47 blockade enables tumor cell phagocytosis by antigen presenting cells (APC), establishing this molecule as a viable therapeutic target in GBM. We additionally investigated the combinatorial effect of CD47 blockade with temozolomide. DNA alkylating agents induce sporadic tumor cell necrosis associated with the extracellular release of damage associated molecular patterns (DAMPs) such as calreticulin, a pro-phagocytic protein whose interaction with low density lipoprotein receptor-related protein 1 (LRP1) facilitates recognition by professional antigen-presenting cells (APCs) and acts as a critical molecular component in promoting immunogenic cell. Our results showed that tumor cell treatment with temozolomide induces plasma membrane expression of calreticulin. Combination therapy resulted in amplified tumor cell phagocytosis and antigen presentation by APC. In addition, preclinical assessment of combination therapy in a syngeneic murine model of GBM resulted in significantly improved survival, characterized by increased intra-tumor penetration of APC cells. These results suggest that the combination of CD47 blockade with temozolomide may enhance tumor immunogenicity, and can improve clinical outcomes demonstrated by mono-therapeutic approaches with conventional chemotherapy. Citation Format: Yaqing Qie, Christina Von Roemeling, Yuanxin Chen, Kevin Shih, Xiujie Liu, Wen Jiang, Joshua Knight, Charles Chan, Irving Weissman, Betty Kim. CD47 blockade with temozolomide can enhance the therapy in glioblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-208. doi:10.1158/1538-7445.AM2017-LB-208

Abstract While growing evidence suggests that tumor microenvironment can modulate stemness and the mesenchymal phenotype, little is known about how these cell states are coordinately regulated. We report that S100A4 controls both stemness and mesenchymal transition in GBM cells. In a spontaneous mouse glioma model and in primary human GBM tumorspheres, S100A4 expression is enriched in tumor-initiating cells and is required for self-renewal and tumor growth in vivo. In addition, greater than 80 percent of S100A4+ cells are associated with the tumor vasculature, a known niche for glioma stem cells, and S100A4 promotes pro-angiogenic factor expression. Interestingly, acidosis induces expression of S100A4 and the EMT regulators SNAIL1/2 and acidosis-induced EMT regulator expression requires S100A4 function. Importantly, S100A4 functions as a molecular switch between the proneural and mesenchymal states in GBM as S100A4 knockdown results in down-regulation of the mesenchymal signature genes, including the “master transcriptional regulators of mesenchymal transition”, and concurrent up-regulation of proneural signature genes. Together, these results indicate that S100A4 sits in a critical node in the molecular network that controls both glioma stem cells and the mesenchymal phenotype. Citation Format: Kin-Hoe Chow, Hee Jung Park, Joshy George, Yuanxin Chen, Andrew Gallup, Wen Jiang, Betty Y. Kim, Kyuson Yun. Regulation of GBM stemness and mesenchymal transition [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4783. doi:10.1158/1538-7445.AM2017-4783

Abstract Mutant peptides presented in cancer are superior vaccine candidates than self peptides. The efficacy of mutant K-Ras, P53 and EFGR peptides have been tested as cancer vaccines in pancreatic cancer, colorectal cancer, and lung adenocarcinoma. However, the occurrence of these mutations in cancer is limited. Further more, these mutant peptides can only be presented by certain MHC alleles. Neoantigens expressed by passenger mutations which are not involved in common proliferative molecular pathways are potential candidates for personalized vaccine design. Such passenger mutations have drawn attention by the recent clinical data that colorectal cancer with a large number of somatic mutations due to mismatch-repair defects are more susceptible to immune checkpoint blockade by PD1 antibody therapy. In this study, we analyzed the next generation sequencing data of 147 lung adenocarcinoma patients from The Cancer Genome Atlas and predicted neoantigens presented by MHC Class I and Class II molecules. We found 18175 expressed clonal somatic mutations, with an average of 124 per patient. Based on such somatic mutations, we predicted 8731 neo-peptides, among which 3888 strong-binders and 4843 weak-binders to MHC Class I molecules were found by NetMHCPan. For HLA-A*02:01, 778 strong binding and 822 weak binding neo-peptides were found. In average, 11 neo-antigen peptides of mutations were predicted per individual patient with HLA-A*02:01 allele. Smokers expressed 5.73 fold higher mutant neo-antigen peptides than non-smokers. The 20 most commonly mutated genes with predicted neo-antigens are TTN, RYR2, MUC16, KRAS, ZFHX4, TP53, USH2A, NAV3, KEAP1, ANK2, DNAH9, ADAMTS12, CSMD1, CSMD3, SPTA1, FAT3, ZNF804A, PAPPA2, PCLO, EGFR. As a positive control, the frequency of known proliferation-related mutations (KRAS, P53, and EGFR) were determined in 147 patients, which are consistent with previously next generation sequencing studies of lung adenocarcinoma. Mutation hotspots were found in proliferation-related mutations (KRAS), but not in passenger mutations. 6950 neo-peptides, including 259 strong binders and 6691 weak binders were found for HLA DRB1, an MHC Class II molecule. INDEL mutation type of neo-epitopes were found in 119 of 147 patients, with an average of 4.34 INDEL mutations presented by MHC Class I and 0.86 INDEL mutations presented by MHC Class II molecule HLA DRB1 in every patient. Our results support the feasibility of discovering individualized mutant peptides as candidates for immunodiagnosis and immunotherapy of lung adenocarcinoma. Neo-epitopes which can be presented by both MHC Class I and MHC Class II molecules are attractive targets for designing long-peptide vaccines. Note: This abstract was not presented at the meeting. Citation Format: Dapeng Zhou, Weijing Cai, Deng Pan, Tan Wen Ling, Jiaqian Wang, Caichun Zhou, Yanyan Lou. Neoantigens predicted by clonal mutation analysis in lung adenocarcinoma patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 628. doi:10.1158/1538-7445.AM2017-628

Abstract Pancreatic cancer (PaCA) is highly refractory to treatment, with a median survival of 6 months. Currently, gemcitabine (Gem), a difluoro analog of deoxycytidine, is the standard care for PaCA, but it renders only a modest survival benefit. Combination therapy, integrating novel agents into standard-of-care regimens, may maximize efficacy of Gem-based therapy. Fibroblast growth factor receptor (FGFR) function is aberrant in pancreatic cancer, and suppression of FGFRs inhibits pancreatic cancer growth and invasion. However, the mechanisms of FGFR action and their therapeutic utility in pancreatic cancer remain to be elucidated. Here we tested the hypothesis that FGFRs can impact Gem sensitivity of PaCA cells by employing pan-FGFR inhibitors and shRNAs against FGFRs. NVP-BGJ398 and AZD4547 are novel FGFR inhibitors (FGFRIs) specific to FGFR1, FGFR2 and FGFR3, and are in various stages of clinical trial. The FGFRIs modestly but significantly inhibited proliferation, migration, and invasion of PANC-1, MIAPaCa-2 cells in wound-healing and trans-well assays. Treatment with low-concentration Gem (7.5nM) enhanced MIAPaCa-2 cell migration at 24hrs, but migration was reduced subsequently, suggesting a sequence of delayed effects of Gem upon cell migration. After 72hrs of exposure, Gem combined with FGFRIs reduced MIAPaCa-2 cell proliferation significantly compared to controls or Gem- and FGFRI alone. Pharmacodynamic modeling was employed to evaluate drug interaction, and the interaction coefficient Psi was estimated by simultaneous modeling of 16 different drug combinations over a period of 0-96 hrs. The analysis estimated Psi=0.78, which indicates synergism. Compared to BGJ398 or Gem alone, the combination also increased the percentage of cells arrested at S phase for a longer period of time. Western blots revealed significant downregulation of ribonucleoside-diphosphate reductase large subunit (RRM1), DNA excision repair protein (ERCC1), and Epithelial-to-Mesenchymal transition biomarkers (vimentin and snail) with FGFRI/Gem treatment compared to Gem-treated cells, indicating that FGFRIs inhibit numerous factors that may contribute to Gem resistance. Loss-of-function by shRNA-mediated knockdown of FGFR1 in MIAPaCa-2 cells showed decreased expression of RRM1, whereas knockdown of FGFR2 and FGFR3 mediated an increase in RRM1 expression, implicating FGFR1 suppression in improving Gem-sensitivity. These results suggest that inhibition of FGFR1 could potentially enhance sensitivity of PaCA to Gem therapy, and that combining FGFRs with gemcitabine is a promising treatment strategy in pancreatic cancer. Citation Format: Qingxiang Lin, Tista Roy Chaudhuri, Ninfa L. Straubinger, Wen Wee Ma, Robert M. Straubinger. FGFR inhibitors enhance gemcitabine sensitivity in pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4072. doi:10.1158/1538-7445.AM2017-4072

Abstract The majority of Pancreatic cancer (PaCA) patients present with metastatic disease and receive little benefit from chemotherapy. Although nanoparticulate carriers can potentially deposit sustained and high doses of chemotherapeutics in tumors, PaCA tumors are poorly perfused, which limits drug deposition. Ten days of oral dosing with NVP-LDE225 (40 mg/kg), an inhibitor of the sonic hedgehog (SHH) pathway (sHHI), increased tumor deposition of blood-borne, fluorescently labeled, sterically-stabilized lipidic (SSL) nanoparticles by enhancing the perfusion and permeability of the otherwise-impermeable microvasculature of PaCA patient-derived xenograft (PDX) tumors. However, literature indicates sHH ablation is associated with increased Epithelial-to-Mesenchymal (EMT) transition and invasiveness in PaCA. Here we investigated the compensatory role of FGFR1 in sHHI-induced tumor progression. Four to 8 daily treatments with the sHHI resulted in no significant reduction of tumor volume, but markedly increased the ratio of tumor cells (human mitochondria+) to Collagen1 in the stroma, in two s.c. low-passage PDX models of varying stromal content. Immunostaining of sHHI-treated tumors revealed no alteration in total FGFR1 expression, but showed a significant elevation in the (1) percent of FGFR1+ pixels correlating with DAPI+ nuclei in tumor (human mitochondria+) cells and stromal cells, (2) number of vimentin+ (EMT) cells, and (3) proliferative index marked by Ki67+ nuclei, compared to controls, suggesting accelerated tumor progression. Nuclear FGFR1 was associated with vimentin+ cells, but was excluded from the nuclei of Ki67+ cells in all groups, implicating nuclear translocation of FGFR1 in EMT. NVP-BGJ398, a small-molecule pan-FGFR inhibitor (FGFRI), administered p.o. for 3 days (15 mg/kg/day) after 7d of sHHI-pretreatment (1) significantly reduced nuclear localization of FGFR1 compared to sHHI-treated animals, (2) reversed the ratio of human mitochondria+ tumor cells to Collagen 1+ stroma, (3) depleted vimentin+ structures by >2 fold, and (4) reduced Ki67+ cells to control levels. FGFRI without sHHI-pretreatment resulted in decreased nuclear FGFR1 relative to controls, but did not impact proliferation, EMT, or tumor cell-to-stroma ratio. Finally, the sHHI/FGFRI sequence increased functional (FITC-L. esculentum lectin+) tumor vessel density and area the perfused by vascular permeability probes (fluorescent SSL) compared to both controls and sHHI-treated animals. FGFRI administration in vehicle-treated controls had no impact on vascular permeability or perfusion. We conclude that the combination of sHHI in sequence with FGFRI may reverse potentially deleterious effects of sHHI and mediate a transient enhancement of tumor permeability, and increase deposition and penetration of nanoparticulate SSL drug formulations into pancreatic cancers. Citation Format: Tista Roy Chaudhuri, Qingxiang Lin, Ninfa L. Straubinger, Wen Wee Ma, Robert M. Straubinger. FGFR1 inhibitor reverses Smoothened inhibition-induced epithelial-to-mesenchymal transition in pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4071. doi:10.1158/1538-7445.AM2017-4071