bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 962: Testing T cell co-potentiation as an antitumor therapeutic strategy in humanized mouse models
Authors: Nelson Alfreda D. , Elsbernd Laura , Schrum Adam G. , Pages Diana Gil
Published: August 2018

We recently described the concept of T cell co-potentiation as means to target antigen recognition by T cells to achieve antitumor immunotherapy in mouse models. Normally, T cell antigen receptor (TCR) engagement by a strong immunogenic antigen induces a CD3 conformational change that exposes a proline-rich sequence (PRS) in the cytosolic domain of CD3 epsilon (CD3e) to dock cytosolic proteins and contribute to the assembly of the signaling cascade that activates T cell immune function. While induction of CD3 conformational change is required by T cells to achieve productive TCR/CD3 signaling, this molecular event fails to occur when the TCR interacts with poorly immunogenic antigens, like most tumor-associated antigens (TAAs). We found that CD3 conformational change can be exogenously induced on T cells using monovalent Fab fragments (Mono-Fabs) specific for the CD3 complex. Anti-CD3 Mono-Fabs expose the CD3e PRS, but fail to crosslink the TCR/CD3, turning them incompetent to trigger T cell activation in the absence of additional stimulation. However, if TCR has bound a weak antigen, one that fails to induce CD3 conformational change intrinsically, then anti-CD3 Mono-Fabs enable TCR/CD3 signaling, and as a result co-potentiate T cell responses to weak antigenic stimulation. Using a mouse model for melanoma (B6 mice and B16-F10 melanoma cell line), we found that treatment of mice with anti-CD3e Mono-Fabs improved antitumor T cell responses against melanoma in an antigen-dependent manner, and dramatically improved the outcome of complementary T cell immunotherapies. Based on these observations, we hypothesize that T cell co-potentiation using anti-human CD3 Mono-Fabs can be exploited as a novel immunotherapy for cancer patients. We have recently identified one anti-human CD3 Mono-Fab that is capable to induce CD3 conformational change in human T cells and allows the co-potentiation of T cell response to weak antigenic stimulation. Here we present data from human T cells and humanized mouse models using this Mono-Fab that support the potential therapeutic value of exploiting T cell co-potentiation to treat cancer patients.

Citation Format: Alfreda D. Nelson, Laura Elsbernd, Adam G. Schrum, Diana Gil Pages. Testing T cell co-potentiation as an antitumor therapeutic strategy in humanized mouse models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 962.
Abstract 989: Organoid cultures from normal and cancer-prone human breast tissues preserve complex epithelial lineages and can form chimeric mammary glands in vivo
Authors: Rosenbluth Jennifer M. , Schackmann Ronald , Li Carman , Sachs Norman , Dillon Deborah , Richardson Andrea , Brock Jane , Garber Judy , Gray Gary Kenneth , Zoeller Jason , Boedicker Mackenzie , Kuiken Hendrick Johannes , Clevers Hans , Brugge Joan
Published: August 2018

Use of cell culture models to investigate the development of cancer associated with familial breast cancer syndromes is challenging because many culture methodologies do not maintain all of the lineages that are present in vivo, particularly the progenitor cells that may be the specific cancer cell-of-origin. We evaluated the ability of organoid culture technology to preserve stem/progenitor and differentiated cell types via long-term propagation of normal human mammary tissues. Tissue samples were obtained from elective reduction mammoplasties or from prophylactic mastectomies for cancer prevention, and were histologically normal. These were grown using conditions similar to organoid cultures for other epithelial cell types, as has been recently described (Sachs et al., Cell 2018). We found that basal stem and luminal progenitor cells could be cultured long-term, and differentiated in culture to generate mature basal and luminal cell types. Analysis of matched organoid cultures and native tissues by mass cytometry (CyTOF) for 38 markers at single-cell resolution confirmed the presence of multiple mammary epithelial cell types in the organoids, and demonstrated that protein expression patterns of the tissue of origin were largely preserved in culture. In addition, we generated a panel of over 40 mammary organoids derived from patients harboring inherited mutations in the cancer predisposition gene BRCA1 and from unaffected controls. Despite interindividual variability, an expansion in luminal progenitor populations associated with BRCA1 mutation could be detected in cultured organoids. Human mammary organoids heterozygous for BRCA1 could be engrafted into the murine mammary gland, resulting in mixed ductal and acinar structures. These studies indicate that this new model system is well suited for studies of aberrant phenotypes associated with BRCA1 mutation and approaches to prevent cancer development in these genetically predisposed tissues.

Citation Format: Jennifer M. Rosenbluth, Ronald Schackmann, Carman Li, Norman Sachs, Deborah Dillon, Andrea Richardson, Jane Brock, Judy Garber, Gary Kenneth Gray, Jason Zoeller, Mackenzie Boedicker, Hendrick Johannes Kuiken, Hans Clevers, Joan Brugge. Organoid cultures from normal and cancer-prone human breast tissues preserve complex epithelial lineages and can form chimeric mammary glands in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 989.
Abstract CT061: Interim results of a phase Ia clinical trial of a PSA, IL-2, GM-CSF containing prostate cancer therapeutic vaccine in PSA defined biochemical recurrent prostate cancer patients
Authors: Head Jonathan F. , Daniels Gregory A. , McKinney Michelle A. , Ongkeko Weg M. , Sakamoto Kyoko , Wang-Rodriguez Jessica
Published: August 2018

While progress has occurred in both cellular and immune checkpoint therapy, the benefit of therapeutic cancer vaccines has been difficult to demonstrate. Prostate cancer may be an exception with the modest activity of sipuleucel-T. We have developed a potentially lower cost and easy to administer therapeutic vaccine with the combination of tumor antigen (PSA) and biologic adjuvants (IL-2 and GM-CSF). This study is a Phase 1a clinical trial of a PSA/IL-2/GM-CSF vaccine in 20 biochemically recurrent hormone-naïve and hormone-independent prostate cancer patients. Major inclusion criteria include adenocarcinoma of the prostate, rising serum PSA and no measurable disease. Phase 1a examines the rate of serious adverse events (SAEs) and dose limiting adverse events (DLAEs) in an initial course of 6 vaccinations (“induction vaccination”). Twenty patients enrolled and nineteen patients completed all six intradermal injections (one patient received 5 vaccines, missed week 11 vaccine) of the PSA/IL-2/GM-CSF vaccine at weeks 1, 2, 3, 7, 11 and 15. None of the patients vaccinated in the Phase 1a portion had a SAE or DLAE with the most common AE relating to grade 1 injection site reactions. Fifteen of the 18 patients who received vaccines had immune responses to PSA as demonstrated in a lymphocyte blastogenesis assay by week 31. Interestingly, after vaccination 14 of 20 patients had an increase in the doubling time of their serum PSA, suggesting a slowing of the growth of the prostate cancer. Four of 20 patients have progressed (3 PSA progression and 1 radiological progression) at 31 weeks follow-up.

Citation Format: Jonathan F. Head, Gregory A. Daniels, Michelle A. McKinney, Weg M. Ongkeko, Kyoko Sakamoto, Jessica Wang-Rodriguez. Interim results of a phase Ia clinical trial of a PSA, IL-2, GM-CSF containing prostate cancer therapeutic vaccine in PSA defined biochemical recurrent prostate cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr CT061.
Abstract CT082: TTFields concurrent with standard of care for the treatment of stage 4 non-small cell lung cancer (NSCLC) following platinum failure: phase III LUNAR study
Authors: Weinberg Uri , Farber Ori , Giladi Moshe , Bomzon Zeev , Kirson Eilon
Published: August 2018

Tumor Treating Fields (TTFields) are a non-invasive, anti-mitotic treatment modality. TTFields disrupt the formation of the mitotic spindle, and dislocation of intracellular constituents. TTFields significantly extend the survival of newly diagnosed glioblastoma patients when combined with temozolomide. Efficacy of TTFields in NSCLC has been shown preclinically and their safety in a phase I/II pilot study with pemetrexed. We hypothesize that adding TTFields to immune checkpoint inhibitor or docetaxel following platinum doublet failure will increase OS.

Methods: Patients (N=534) with squamous or non-squamous NSCLC are enrolled in the LUNAR phase III study [NCT02973789]. Patients are stratified by their selected standard therapy (immune checkpoint inhibitors or docetaxel), histology (squamous vs. non-squamous) and geographical region. Key inclusion criteria are disease progression while on or after platinum-based systemic therapy, ECOG 0-2, no electronic medical devices in the upper torso, and absence of brain metastasis. Docetaxel or immune checkpoint inhibitors are given at standard doses. TTFields are applied to the upper torso for 18 hours/day, allowing patients to maintain daily activities. TTFields are continued until progression in the thorax and/or liver. Follow up is performed every 6 weeks, including CT scans of the chest and abdomen. On progression in the thorax and/or liver, patients have three post-progression follow up visits and are later followed monthly for survival. The primary endpoint is superiority in OS between patients treated with TTFields in combination with the standard of care treatments, compared to standard of care treatments alone. Key secondary endpoints compare the OS in patients treated with TTFields and docetaxel Vs. those treated with docetaxel alone, and patients treated with TTFields and immune checkpoint inhibitors Vs. those treated with immune checkpoint inhibitors alone. An exploratory analysis will test non-inferiority of TTFields with docetaxel compared to checkpoint inhibitors alone. Secondary endpoints include progression-free survival, radiological response rate, quality of life based on the EORTC QLQ C30 questionnaire and severity & frequency of adverse events. The sample size is powered to detect a HR of 0.75 in TTFields-treated patients versus control group.

Citation Format: Uri Weinberg, Ori Farber, Moshe Giladi, Zeev Bomzon, Eilon Kirson. TTFields concurrent with standard of care for the treatment of stage 4 non-small cell lung cancer (NSCLC) following platinum failure: phase III LUNAR study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr CT082.
Abstract CT085: Randomized phase II trial of hydroxychloroquine in combination with gemcitabine/nab-paclitaxel to inhibit autophagy in pancreatic cancer: A SU2C-funded trial
Authors: Karasic Thomas B. , O'Hara Mark H. , Loaiza-Bonilla Arturo , Reiss-Binder Kim A. , Teitelbaum Ursina R. , Borazanci Erkut , Jesus-Acosta Ana M. De , Redlinger Colleen , Burrell Jessica A. , Hoff Daniel D. Von , Laheru Daniel A. , Amaravadi Ravi K. , Drebin Jeffrey A. , O'Dwyer Peter J.
Published: August 2018

Autophagy, a regulated metabolic process that enables energy conservation in adverse environmental circumstances, has been shown in animal models to confer resistance to chemotherapy, which can be reversed by hydroxychloroquine (HCQ). After establishing safety in a Phase I run-in, 112 patients were randomized to receive standard gemcitabine (1000mg/m2) and nab-paclitaxel (125 mg/m2) weekly x 3 every 4 weeks, with (n=55 evaluable) or without (n=55 evaluable) HCQ 600mg bid. Median age was 65 (range 43-86) of whom 47% were aged > 65, 40% were female, 93% Caucasian, ECOG PS 0/1 51%/49%, and did not differ between the arms. Treatment-related side-effects were consistent with prior studies, and were balanced by arm, except for Grade 3/4 neutropenia (42 vs 23%), nausea (9 vs 0%), and fatigue (7 vs 0%), all higher with HCQ. Partial responses were observed in 21/46 (46%) with HCQ, and 8/48 (17%) without HCQ. The primary endpoint, overall survival at 12 months, was 41% (95% CI 27-53%) in the HCQ arm and 51% (95% CI 36-63%) in the non-HCQ arm. Median progression-free survival was 5.7 months (95% CI 4.0-9.3) in the HCQ arm and 6.4 months in the non-HCQ arm (95% CI 4.5-7.6). Median overall survival was 11.1 months (95% CI 9.0-14.2) in the HCQ arm and 14.4 months (95% CI 9.5-15.6) in the non-HCQ arm. Genomic testing was performed on a subset of the patients (41%), in whom outcomes did not differ by p53 mutational status. We conclude that based on the primary endpoint, the addition of the autophagy reversal agent HCQ did not improve the survival of patients with metastatic pancreatic cancer. Response rates in patients who received HCQ were higher, and toxicity was tolerable. Autophagy reversal might be explored in the management of locally-advanced disease.

Citation Format: Thomas B. Karasic, Mark H. O'Hara, Arturo Loaiza-Bonilla, Kim A. Reiss-Binder, Ursina R. Teitelbaum, Erkut Borazanci, Ana M. De Jesus-Acosta, Colleen Redlinger, Jessica A. Burrell, Daniel D. Von Hoff, Daniel A. Laheru, Ravi K. Amaravadi, Jeffrey A. Drebin, Peter J. O'Dwyer. Randomized phase II trial of hydroxychloroquine in combination with gemcitabine/nab-paclitaxel to inhibit autophagy in pancreatic cancer: A SU2C-funded trial [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr CT085.
Abstract CT089: The Dutch Oncology Research Platform (DORP), a national infrastructure for investigator-initiated multicenter clinical cancer research
Authors: Punt Cornelis J.
Published: August 2018

Background Investigator-initiated clinical cancer research (IICCR) contributes in a major way to improved prognosis of cancer patients. However, increased complexity of legislation and regulation, lack of uniformity in local procedures among participating hospitals, and increasing demands on monitoring seriously endanger IICCR, since insufficient staff/funding is available to meet these requirements. In addition, better characterization of patient subgroups requiring specific treatments hamper the feasibility of randomized studies with classic design, and calls for innovative study designs. In The Netherlands, tumor-specific research groups exist for almost every tumor type, which however have different levels of professionalization, but which all face the same problems. This seriously hampers future development of IICCR.

Methods A national infrastructure (DORP) has been developed to facilitate IICCR. DORP has 3 pillars: statistical support, monitoring, and project management. DORP provides statistical and logistic support in protocol development according to up-to-date standards, and, once financial support for the conduct of the study (i.e. datamanagement) has been obtained, provides support for monitoring and project management. The latter includes implementation of the study in participating hospitals, and administrative/financial management of the study. DORP statisticians will develop innovative trial designs that provide more feasible alternatives to classic phase 3 studies in small subgroups of patients, and DORP project managers will aim to harmonize local study procedures in hospitals in order to speed up study initiation.

Results The DORP initiative has been received with great enthousiasm among stakeholders (i.e. research groups, patient advocacy groups, large trial offices, professional and hospital organisations, insurance companies). DORP has recently received a 6.5 million euro starting grant from the Dutch Cancer Society (KWF Kankerbestrijding), which has allowed to established DORP as a cooperation in which all stakeholders participate. A total of approx. 30 statisticians, monitors and project managers will be recruited and will be employed at institutions with established expertise in the respective fields. A director has been commissioned by the board. Trial development with DORP support has been initiated in January 2018.

Conclusion The objective of DORP is more trials, for more patients, of better quality, that are timely completed. DORP may internationally provide an example for IICCR support.

Citation Format: Cornelis J. Punt. The Dutch Oncology Research Platform (DORP), a national infrastructure for investigator-initiated multicenter clinical cancer research [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr CT089.
Abstract CT139: Durability of responses to the combination of SD-101 and pembrolizumab in advanced metastatic melanoma: Results of a phase Ib, multicenter study
Authors: Ribas Antoni , Medina Theresa , Kummar Shivaani , Amin Asim , Drabick Joseph J. , Barve Minal , Daniels Gregory , Wong Deborah L. , Schmidt Emmett V. , Leung Abraham C. , Janssen Robert
Published: August 2018

Background: This phase 1b trial evaluates the safety and antitumor efficacy of the combination of SD-101, a synthetic CpG-oligonucleotide that stimulates Toll-like receptor 9 (TLR9), and pembrolizumab in patients with recurrent Stage IIIC/IV malignant melanoma.

Methods: Dose escalation of SD-101 used a modified 3+3 design. SD-101 was injected in a single tumor lesion (weekly x 4 doses then every 3 weeks x 7 doses over 6 months) at 1, 2, 4, or 8 mg and pembrolizumab was administered intravenously at 200 mg every 3 weeks up to 2 years. Tumor responses were assessed per investigator using RECIST v1.1 taking into account both injected and non-injected lesions.

Results: Of the 22 patients, 9 were naïve to anti-PD-1/L1 therapy at baseline and 13 had progressive disease while receiving prior anti-PD-1/L1 therapy. Treatment was well tolerated with no Grade 3 or higher treatment-related AEs in longer term follow up. Among the 9 patients who were anti-PD-1/L1 naïve, best objective responses were CR: 2, PR: 5, PD: 1, not evaluated [NE]: 1. Median PFS, duration of response, and OS have not been reached. Estimated 12 month PFS was 88% and OS was 89%. After a median of 18 months of follow-up, 86% of responses were ongoing. One patient with a PR developed progressive disease after 20 months of treatment. Among patients who had received prior anti-PD-1/L1 therapy, best objective responses were PR: 2, SD: 5, PD: 5, NE: 1. One patient with stable disease and 1 patient with a PR continued on combination therapy without progression for at least 10.5 months. The other 10 patients developed progressive disease ranging from 1.5 to 8 months after enrollment. Percent change in tumor size from baseline in injected and non-injected lesions will also be presented.

Conclusion: These early results suggest that combining an intratumoral TLR9 innate immune stimulant with PD-1 blockade can potentially provide more clinical responses that are meaningfully durable than PD-1 blockade alone based on historical data.

Citation Format: Antoni Ribas, Theresa Medina, Shivaani Kummar, Asim Amin, Joseph J. Drabick, Minal Barve, Gregory Daniels, Deborah L. Wong, Emmett V. Schmidt, Abraham C. Leung, Robert Janssen. Durability of responses to the combination of SD-101 and pembrolizumab in advanced metastatic melanoma: Results of a phase Ib, multicenter study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr CT139.
Abstract CT159: Olaparib combinations (OLAPCO)for homology-directed DNA repairdefects (HDR)
Authors: Eder Joseph P. , Shapiro Geoffrey I. , Cleary James M. , Keedy Vickie L. , Keedy Vickie L. , Bindra Ranjit , Shuch Brian , Sklar Jeffrey , Do Khanh , Juergensmeier Juliane
Published: August 2018

INTRODUCTION: Genetic profiling of tumors used to identify treatment choices for cancer patients focus on kinase mutations and well-defined translocations; multiple clinical trials have been initiated for specific kinase-targeted agents. About 15-20% of patients have actionable mutations. HDR have been found in ~ 20 % of patient tumors molecularly profiled at the Yale Cancer Center. Mutations in HDR related genes often occur in addition to other mutations in signaling pathways. Specific targeted treatment options for these patients are limited, as breast and ovarian cancer mutations are the only indication for the HDR pathway targeted PARP inhibitors. Additional drugs are in development, but few clinical studies based on HDR other than BRCA1/2 mutations are open.

METHODS: The OLAPCO study targets HDR in patients irrespective of tumor origin. The protocol includes four arms to which patients can be enrolled based on tumor profiling results: olaparib monotherapy (PARP inhibitor) for HDR; olaparib & AZD5363 ( AKT inhibitor) for PIK3CA/AKT/PTEN pathway dysregulation and ARID1A mutations; olaparib & AZD1775 ( wee1 inhibitor) for TP53 mutations and olaparib & AZD6738 (ATR inhibitor) for HDR and olaparib pre-treated patients. The clinical endpoints are to determine tumor overall response rate (ORR) and clinical benefit rate (CBR) by RECIST after 16 weeks of treatment.

RESULTS: Arms a and b are open to patient accrual. Together, 34 pts have been treated, 12 remain on treatment, with no new or > Gr 2 (CTCAE4) treatment-related toxicities.

TABLE 1Olaparib total = 23; median # cycles= 2, range 1-10+ ; 7 ongoingMutationATMBRCA1IDH1/2FHPALB2CHK2BAP1ARID 1A#52821212ORR1 PRCBR10201000TABLE 2Olaparib & AZD 5363 total = 12; median # cycles = 2, range 1-8+; 5 ongoingMutationAKT1AKT3AKT2 & PALB2ARID 1AARID 1A & PIK3CAPTEN#321411ORR1CBR10101

DISCUSSION: HDR defects are actionable mutations, increasing the number of patients for whom tumor profiling can identify a therapeutic option. This exploratory trial is expanding the therapeutic spectrum of olaparib alone and in combination, to include heretofore non-canonical alterations in HDR, beyond BRCA1/2. This trial design allows additional molecular targets, agents & combinations to be rapidly evaluated.

AstraZeneca support is gratefully acknowledged.

Citation Format: Joseph P. Eder, Geoffrey I. Shapiro, James M. Cleary, Vickie L. Keedy, Vickie L. Keedy, Ranjit Bindra, Brian Shuch, Jeffrey Sklar, Khanh Do, Juliane Juergensmeier. Olaparib combinations (OLAPCO)for homology-directed DNA repairdefects (HDR) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr CT159.
Abstract LB-005: Anti-tumoral effects of novel lymphatic delivery of anti-CTLA-4 in a metastatic murine breast cancer model
Authors: Kwon Sunkuk , Velasquez Fred Christian , Ross Russel F. , Hwu Wen-Jen , Sevick-Muraca Eva M.
Published: August 2018

The cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) is an immune checkpoint receptor expressed by the activated T-cells and plays a key role in modulating T-cell function. Blockade of CTLA-4 after systemic administration can induce an antitumor immune response by promoting the activation and proliferation of tumor-specific T cells. Despite important clinical benefits, systemic intravenous administration of anti-CTLA-4 antibody often results in off-target toxicities (i.e., severe immune-related adverse events) by inappropriately activating naïve-T cells in distant lymph nodes (LNs) and other lymphoid tissues where non-tumor, self-antigens rather than tumor antigens are presented. Thus, efficient direct delivery of checkpoint inhibitors into the regional lymphatics could provide maximal drug exposure to targets within tumor-draining LN basins and their efferent lymphatic vessels before emptying into the blood vasculature for systemic delivery to the tumor. In this study, we used a novel, infusion device, SOFUSATM, consisting of a 66 mm2 microneedle array with a 6 micron polyether ketone nanotopographical film heat-formed over each microneedle to infuse therapeutic doses into the lymphatics in mice. 4T1 mouse mammary tumor cells transfected with luciferase (4T1-luc) were injected into the mammary fat pad of BALB/C mice. Anti-(cytotoxic T-lymphocyte-associated protein-4 (CTLA-4) antibody (10mg/kg) was administered on days 11, 15, 19, and 23 via SOFUSATM. SOFUSATM infusion occurred on the right dorsal back. While further studies are underway, our preliminary data demonstrated that when the repeated dose of anti-mCTLA-4 was successfully delivered via SOFUSATM, significant tumor shrinkage occurred when compared to the untreated group and the group of mice treated with conventional intraperitoneal (i.p.) administrations. Our bioluminescence imaging data showed no lymph node and distant organ metastases in mice dosed with SOFUSATM as compared to the untreated mice and mice with i.p. treatment, both of which showed LNs, lung, or liver metastases. Our study may revolutionize immunotherapies by moving from advanced, systemic to “first-line” loco-regional, cancer treatment.

Citation Format: Sunkuk Kwon, Fred Christian Velasquez, Russel F. Ross, Wen-Jen Hwu, Eva M. Sevick-Muraca. Anti-tumoral effects of novel lymphatic delivery of anti-CTLA-4 in a metastatic murine breast cancer model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-005.
Abstract LB-015: Differential therapeutic responses of MET oncogenic activations to Met kinase inhibitor and neutralizing antibody
Authors: Kou Jianqun , Staal Ben , Musich Phillip R. , Kang Liang , Qin Yuan , Yao Zhi Q. , Zhang Boheng , Angela Tam , Huang Alan , Hao Huaixiang , Woude George F. Vande , Xie Qian
Published: August 2018

Purpose: MET inhibitors are in clinical trials against several cancer types, but the mechanisms toward vulnerability remain elusive. Here we characterized the molecular basis of MET amplification (METamp) and HGF-autocrine driven tumors in response to MET tyrosine kinase inhibitors (TKI) and neutralizing antibodies.

Experimental Design: METamp (MKN45 and MHCC97H) and HGF-autocrine activation (JHH5 and U87) cells were treated by the MET kinase inhibitor (INC280) and the anti-MET monoclonal antibody (MetMab) to determine the sensitivity and biological responses in vitro. Tumor inhibition was evaluated in vivo using SCID and SCIDhgf mouse models, respectively. HGF-mediated angiogenesis was measured by using the human endothelial cells (HUVEC) tube formation assay.

Results: MKN45 and MHCC97H cells are more sensitive than JHH5 and U87 cells to INC280 treatment but are unresponsive to MetMab. In METamp cells, INC280 induced a DNA damage response with activation of repair through the p53BP1/ATM signaling pathway. Although INC280 and MetMab showed a moderate inhibitory effect on - JHH5 and U87 cells in vitro, both treatments potently suppressed tumor growth in mouse models. We found that HGF stimulation promotes human HUVEC cell tube formation via the Src pathway. INC280 or MetMab inhibited tube formation; thus in HGF-autocrine tumors, the endothelial cells are the secondary targets of tumor-derived HGF and MET inhibition.

Conclusion: METamp and HGF-autocrine activation favor different molecular mechanisms, such as a DNA damage response or angiogenesis. Because individual types of MET oncogenic activation may respond to MET inhibitors differently, combination strategies should be developed based upon the molecular subtypes of the tumors.

Citation Format: Jianqun Kou, Ben Staal, Phillip R. Musich, Liang Kang, Yuan Qin, Zhi Q. Yao, Boheng Zhang, Tam Angela, Alan Huang, Huaixiang Hao, George F. Vande Woude, Qian Xie. Differential therapeutic responses of MET oncogenic activations to Met kinase inhibitor and neutralizing antibody [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-015.
Abstract LB-026: iDEP: integrated differential expression and pathway analysis of RNA-Seq data
Authors: Ge Steven X.
Published: August 2018

Analysis and interpretation of RNA-Seq data remain a challenge for bench scientists. We aim to develop a user-friendly web application for exploratory data analysis (EDA), differential expression, and pathway analysis using summarized gene-level data from RNA-Seq, DNA microarray or other platforms. The key idea of iDEP (integrated Differential Expression and Pathway analysis) is to make many powerful R/Bioconductor packages easily accessible by wrapping them under a graphical interface, alongside annotation databases. For EDA, it performs hierarchical clustering, k-means clustering, and principal component analysis. iDEP detects differentially expressed genes using the limma and DESeq2 packages. For a group of co-expressed genes, it identifies enriched gene ontology (GO) terms as well as transcription factor binding motifs in promoters. Pathway analysis can be performed using packages like GAGE, GSEA, PGSEA, or ReactomePA. iDEP can also detect chromosomal gain or loss using the PREDA package. iDEP uses annotation of 163 metazoa and 45 plant genomes in Ensembl for ID mapping and GO functional categorization. Common gene IDs including microarray probe names can be automatically recognized. Pathway information was also compiled from databases like KEGG, Reactome, MSigDB, GSKB, GeneSetDB, and araPath. As an example, we analyzed an RNA-Seq dataset involving siRNA-mediated Hoxa1 knockdown in lung fibroblasts, and identified the downregulation of cell-cycle genes, in agreement with previous studies. Our analyses also reveal the possible roles of E2F1 and its target genes, including microRNAs, in blocking G1/S transition, and the upregulation of genes related to cytokines, lysosome, and neuronal parts. iDEP enables users to conduct in-depth bioinformatics analysis of transcriptomic data through a graphical interface. Freely available at http://gelab.org/idep/

Citation Format: Steven X. Ge. iDEP: integrated differential expression and pathway analysis of RNA-Seq data [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-026.
Abstract LB-028: Deep genomic, immune-infiltrate and neoantigen-based stratification of cancer predicts response to immune checkpoint inhibition
Authors: Ronen Roy , Kluge Boguslaw , Dutkowski Janusz
Published: August 2018

Background: Immunotherapies are revolutionizing cancer care but identifying patients which are most likely to achieve clinical benefit remains a key challenge. Available predictors such as PD-1/PD-L1 expression, tumor mutation burden or MSI-H/d-MMR status have shown promise in predicting response in specific cancer types, they fail to explain or predict response for many patients. A more comprehensive understating of biology underlying response to therapy is needed.

Approach: We developed an integrated machine-learning system that uses a mathematical model of cancer subtypes and a neoantigen fitness model to stratify multimodal omics data along with associated profiles of immune infiltration and immune pathway activity. The system holistically determines the underlying pathway-level architecture of genomic and other molecular alterations in each cancer. It further uses a new immune pathway deconvolution approach to determine subtype-specific immune-infiltrates and activation of key immune and immune-modulating pathways that may be associated with response to therapy.

Results: The system is applied to stratify and interrogate subtypes in over 40 cancer types. Here we discuss results for two of the histologies for which immunotherapies are being pursued clinically. Comprehensive systems-level interrogation of tumor genomic architecture and tumor microenvironment leads to new molecular sub-typings for colorectal adenocarcinoma and skin cutaneous melanoma, here referred to as COADREAD_MUT_CNV_MODEL_6 and SKCM_MUT_CNV_MODEL_3, respectively. The model-based subtypes uncover critical patterns of co-occurring and mutually-exclusive somatic alterations not apparent using previous approaches, and show strong correlation with immune infiltration. Further integration of the subtyping model with a new neoantigen fitness model significantly improves performance in predicting response to checkpoint inhibition compared to existing methods.

Citation Format: Roy Ronen, Boguslaw Kluge, Janusz Dutkowski. Deep genomic, immune-infiltrate and neoantigen-based stratification of cancer predicts response to immune checkpoint inhibition [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-028.
Abstract LB-032: Magnetic 3D bioprinting for the identification of tumor-specific therapies from PDX-derived 3D cultures
Authors: Souza Glauco , Bartholomeusz Geoffrey
Published: August 2018

Great efforts have been undertaken in the field of cancer to develop clinically relevant pre-clinical models to permit the translation of research advances into agents that can improve patient outcomes. The use of 2D monolayer and 3D multicellular tumor spheroid cell culture systems has made this process remarkably costly and inefficient. This outcome is a result of discrepancies between simplified in-vitro 2D cell culture models used in drug screening platforms and the complex 3D tumor microenvironment targeted by these same drugs. Patient-derived xenograft (PDX), in which patient tumor tissue implanted into immune-deficient mice, provide improved models of human tumor biology relative to immortalized cancer cell lines. Such models offer possibilities for better preclinical testing of new therapies for the treatment of cancer. A drawback associated with the use of PDX systems is its requirement for immense resources and animals. Also, animal models have limited relevance as a platform for high throughput drug screening. To overcome such limitations, we applied magnetic 3D bioprinting to develop a PDX-derived ex-vivo tumor tissue platform for high-throughput drug screening. The PDX-derived tissue generated using magnetic 3D bioprinting mirrors the original PDX tumor in both tissue architecture and genetic signature. We confirm the predictive value of our screening template by demonstrating a similar response outcome between our model and a PDX mouse system generated from the same original tumor tissue and treated with the same panel of drugs. Utilizing an Inflammatory Breast Cancer (IBC) PDX system as our model system, we screened our ex-vivo tumor tissue template with the anti-cancer drug library and identified 7 lead candidates that had a stronger anti-tumor activity compared to the standard of care agents used to treat IBC patients in the clinics. Incorporating our ex-vivo tumor tissue high throughput screening platform as an early step in a PDX system will permit the identification of new and effective tumor-specific therapies at a time, cost, and resource efficient manner.

Citation Format: Glauco Souza, Geoffrey Bartholomeusz. Magnetic 3D bioprinting for the identification of tumor-specific therapies from PDX-derived 3D cultures [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-032.
Abstract LB-048: c-Kit enhances ovarian cancer stemness via upregulating phospho-PHB in the lipid raft domain of the plasma membrane
Authors: Liang Shu-Mei Y. , Fang Chia-Hsun , Chang Pu-Yuan , Liang Chi-Ming
Published: August 2018

Phosphorylation of prohibitin (PHB) at threonine 258 (phospho-PHBT258) and tyrosine 259 (phospho-PHBY259) in human cancer-cell membranes is associated with an increase in the invasiveness and metastasis of cancer cells. Phospho-PHBY259 can regulate phospho-PHBT258 but not vice versa. Here, we found that stably overexpressing c-Kit in SKOV3 ovarian cancer cells (SKOV3_c-Kit) increased phospho-PHBY259, phospho-PHBT258 and PHB levels in the lipid raft domain of the plasma membrane. On the other hand, inhibition of c-Kit expression decreased PHB expression in SKOV3_c-Kit and Kuramochi ovarian cancer cells. Immunoprecipitation analysis showed that c-Kit interacts with PHB. SKOV3_c-Kit cells or SKOV3 cells overexpressing PHB in the lipid raft had higher levels of Notch3, NICD3 (Notch3 intracellular domain) and ABCG2 protein expression as compared to wild-type SKOV3 cells. Moreover, generation of SKOV3-G4 cells through serial passage of SKOV3 cells into nude mice ovary 4 times, revealed that the levels of c-Kit, Notch3, NICD3, phospho-PHBY259, phospho-PHBT258 and PHB in the lipid raft domain of SKOV3-G4 were significantly higher than wild-type SKOV3 cells. The SKOV3-G4 cells formed spheroids with stem cell-like properties such as higher expression of stem cell markers (Oct4, Nanog, SOX2). In addition, SKOV3-G4 cells exhibited more clonogenic potential and migration/invasion ability than wild-type SKOV3. These results suggest that c-Kit interacts with PHB in the lipid raft domain to phosphorylate PHB to increase invasiveness and stemness in ovarian cancer.

Citation Format: Shu-Mei Y. Liang, Chia-Hsun Fang, Pu-Yuan Chang, Chi-Ming Liang. c-Kit enhances ovarian cancer stemness via upregulating phospho-PHB in the lipid raft domain of the plasma membrane [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-048.
Abstract LB-065: An enzyme-directed pro-immunotherapeutic activated by muliti-drug resistance
Authors: Nielsen Amy E. , Mancini Rock J.
Published: August 2018

Synthetic agonists of innate immune cells are of interest to immunologists due to their synthesis from well-defined materials, optimized activity, and monodisperse chemical purity. Recently we have reported the approach of Bystander-Assisted ImmunoTherapy (BAIT) that exploits the phenomenon of drug efflux in multi-drug-resistant cancers to elicit a localized anti-cancer immune response. In this context we examine the imidazoquinoline class of immunostimulants for susceptibility to drug efflux and begin to determine structural characteristics that drive efflux susceptibility for the Toll-Like Receptor agonist family of immmunsotimulants.

Citation Format: Amy E. Nielsen, Rock J. Mancini. An enzyme-directed pro-immunotherapeutic activated by muliti-drug resistance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-065.
Abstract LB-117: Role of ectoenzymes CD39 and CD73 in the immune response to multiple myeloma
Authors: Yang Rui , Elsaadi Samah , Misund Kristine , Slupphaug Geir , Menu Eline , Hay Carl , Cooper Zac , Vanderkerken Karin , Børset Magne , Sponaas Anne Marit
Published: August 2018

Multiple Myeloma is a cancer of the plasma cells in the bone marrow. Despite from the fact that new drugs have increased survival in the past decade, it is still an incurable cancer. It is therefore important to develop new drugs. Tumor immunosurveillance contributes to the control of cancer growth, but there many different escape mechanisms utilized by tumour to evade anti-tumour immune responses. One of the largely unexplored mechanisms of immune suppression in multiple myeloma is the role of adenosine in the bone marrow microenvironment, which now represents an attractive new therapeutic target for cancer therapy. 2 ectoenzymes, CD39 and CD73 are important for converting extracellular ATP to adenosine. When investigating the role of adenosine in the immune response to multiple myeloma, we found increased concentration of adenosine in the bone marrow plasma from myeloma patients compared with healthy controls. CD39 (ENTPD1) was expressed by myeloma cell lines and on myeloma cells from patients. CD73 was found on leukocytes and stromal cells in the bone marrow. Although very few cells expressed CD73 in the bone marrow, these cells could clearly convert AMP to adenosine. ATP was converted to AMP by CD39+ myeloma cell lines, and CD73+ stromal cells converted AMP to adenosine. Importantly, co-culture of CD39+ myeloma cells and CD73+ stromal cells catalyzed extracellular ATP to immunosuppressive adenosine. Indeed, we found that CD3 T cells stimulated with anti CD3/CD28 beads in the presence of supernatants from co-cultured cells did not proliferate as well as T cell culture without this supernatant. The fact that this proliferation was inhibited by the ZM241385 inhibitor, suggested that the suppression was mediated by the A2AR receptor. The CD39 inhibitor POM1 inhibited adenosine generation in the co-culture system as well as T cell proliferation in vitro. We have recently obtained the CD73 antibody MEDI199447 from MedImmune and are testing the ability of this antibody to prevent adenosine production and T cell proliferation in vitro, as well as the ability to reduce tumor growth in a mouse model of multiple myeloma. The clinical importance of the adenosine pathway in multiple myeloma found when analyzing patient data. Using the publicly available coMMpass database, we found that patients with high expression of CD39 had significantly reduced progression free and overall survival.

Citation Format: Rui Yang, Samah Elsaadi, Kristine Misund, Geir Slupphaug, Eline Menu, Carl Hay, Zac Cooper, Karin Vanderkerken, Magne Børset, Anne Marit Sponaas. Role of ectoenzymes CD39 and CD73 in the immune response to multiple myeloma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-117.
Abstract LB-128: The aryl hydrocarbon receptor as driver of cancer immunity
Authors: Sherr David , Kenison-Whte Jessica , Wang Zhongyan
Published: August 2018

The Aryl Hydrocarbon Receptor (AHR) has been identified as a driver of cancer progression and cancer immunity in the tryptophan-IDO/TDO-kynurenine-AHR pathway. Over the last 10 years, studies from several laboratories have demonstrated that AHR ligand-mediated immunosuppression is effected, at least in part, through AHR regulation of inflammatory or inhibitory T cell subsets and immune suppressive MDSCs. These results suggest that the AHR is a driver of cancer immunity. To test this hypothesis, we used molecular and pharmacological approaches to regulate AHR activity in multiple murine cancer models and then measured multiple immune phenotypes. The resulting data indicate that a novel, non-toxic AHR inhibitor, HP163 (Hercules Pharmaceuticals), reduces tumor growth in orthotopic models of oral (MOC1), breast (4T1), and skin (B16) cancers in immunocompetent hosts. In the MOC1 model, HP163 significantly reduces the number of CD11b+PD-L1+ or CD11b+CCR2+ cells in the tumor draining lymph node. AHR knockdown in MOC1 cells with CRISPR/Cas9 decreases the percentage of CD11b+PD-L1+ tumor infiltrating cells and increases tumor infiltrating CD4+ and CD8+ T cells. This decrease in cells with an immunosuppressive phenotype is accompanied by a complete lack of orthotopic tumor growth. Furthermore, mice having received AHR- MOC 1 cells are resistant to a second challenge with wildtype AHR+ MOC1 cells several months after the primary inoculation. These data suggest that the presence of the AHR in the tumor itself is sufficient to induce immunosuppression. Preliminary data suggest that AHR+ myeloid cells mediate suppression of tumor immunity. These results collectively suggest that the AHR may be targeted for immunotherapy of several cancers and that HP163 may represent a novel approach to cancer therapy.

Citation Format: David Sherr, Jessica Kenison-Whte, Zhongyan Wang. The aryl hydrocarbon receptor as driver of cancer immunity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-128.
Abstract LB-137: Integrated genomics: drug screening and personalized xenograft development approach to identify precision treatments for aggressive pediatric brain tumors
Authors: Tsoli Maria , Wadham Carol , Pinese Mark , Failes Tim , Joshi Swapna , Mould Emily , Yin Julia , Gayevski Velimir , Kumar Amit , Kaplan Warren , Ekert Paul , Franshaw Laura , Gifford Andrew , Weber Martin , Rodriguez Michael , Mayoh Chelsea , Cohn Richard , Arndt Greg , Lock Richard , Tyrrell Vanessa , Norris Murray , Haber Michelle , Lau Loretta , Quang Dong Anh Khuong , Wong Marie , Trahair Toby , Marshall Glenn M. , Cowley Mark J. , Ziegler David S.
Published: August 2018

Brain tumors represent the most common solid tumors in childhood and account for significant morbidity and mortality. The lack of advances in treatment of the brain tumors is hindered by our lack of knowledge about the molecular pathogenesis of these malignancies as well as the availability of brain-permeable therapies. Personalised medicine approaches have the potential to enhance diagnostic, prognostic and / or therapeutic information. The TARGET study is a feasibility pilot study which focused on the development of genomic platforms, high throughput drug screening (HTS), and PDX models in order to identify personalised therapies for patients with aggressive pediatric tumors. In the pilot phase 59 patients were enrolled of which 28 were brain tumor patients (47%). In 28 curated cases molecular analysis identified a reportable somatic anomaly (SNVs, gene fusions, CNVs) in 82% (23/28), of which 78% (18/23) had actionable molecular aberrations. Four cases had a reportable germline predisposition variant and in 1 case, the genomic findings changed the primary diagnosis. Fresh tissue collection permitted in vitro HTS (112 single agents) in 58% of cases to date. Four PDX models have been established with additional 8 currently still being evaluated. This feasibility study demonstrates that an integrated approach based on genomic, in vitro and in vivo drug efficacy testing may be useful to guide the management of aggressive pediatric brain tumors.

Citation Format: Maria Tsoli, Carol Wadham, Mark Pinese, Tim Failes, Swapna Joshi, Emily Mould, Julia Yin, Velimir Gayevski, Amit Kumar, Warren Kaplan, Paul Ekert, Laura Franshaw, Andrew Gifford, Martin Weber, Michael Rodriguez, Chelsea Mayoh, Richard Cohn, Greg Arndt, Richard Lock, Vanessa Tyrrell, Murray Norris, Michelle Haber, Loretta Lau, Dong Anh Khuong Quang, Marie Wong, Toby Trahair, Glenn M. Marshall, Mark J. Cowley, David S. Ziegler. Integrated genomics: drug screening and personalized xenograft development approach to identify precision treatments for aggressive pediatric brain tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-137.