bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 37: AKT1 activation promotes brain metastasis in a mouse model of melanoma
Authors: Kircher David , Cho Joseph , Robinson James , Arave Rowan , Green Russell , Chen Guo , Davies Michael , Grossmann Allie , VanBrocklin Matthew , McMahon Martin , Holmen Sheri
Published: May 2016

Brain metastases are a major cause of melanoma-related mortality and morbidity. The molecular mechanisms driving melanoma to the brain remain largely unclear, adding to the difficulties in both treating and prognosticating risk for this disease complication. One of the great barriers impeding the study of melanoma brain metastasis is the lack of an in vivo model that faithfully mimics the human disease. Based on previous studies implicating aberrant AKT signaling in human melanoma brain metastases, we evaluated the effect of activated AKT1 expression in non-metastatic BRAFV600E/INK4A-ARFNull mouse melanomas in vivo. Expression of activated AKT1 in this context resulted in highly metastatic melanomas with lung and brain metastases in 67% and 17% of our mice, respectively. Surprisingly, silencing of PTEN in BRAFV600E/INK4A-ARFNull melanomas did not yield a statistically significant metastatic phenotype but in combination with activated AKT1 resulted in decreased tumor latency and development of brain metastases in nearly 80% of tumor-bearing mice. Furthermore, we found evidence of increased mTOR signaling in BRAFV600E/INK4A-ARFNull mouse melanomas expressing activated AKT1 relative to PTEN-null counterparts. This study advances the field of melanoma brain metastasis by (1) providing the first in vivo, autochthonous model of melanoma with spontaneous metastasis to the lungs and brain, (2) demonstrating that expression of activated AKT1 is sufficient to elicit brain metastasis in BRAFV600E/INK4A-ARFNull mouse melanomas and (3) revealing distinct differences and cooperation between AKT1 activation and PTEN silencing in metastatic melanoma progression. In summary, this work advances our knowledge of the mechanisms driving melanoma brain metastasis and may provide valuable insights for the clinical management of this disease.

Citation Format: David Kircher, Joseph Cho, James Robinson, Rowan Arave, Russell Green, Guo Chen, Michael Davies, Allie Grossmann, Matthew VanBrocklin, Martin McMahon, Sheri Holmen. AKT1 activation promotes brain metastasis in a mouse model of melanoma. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Integrating Clinical Genomics and Cancer Therapy; Jun 13-16, 2015; Salt Lake City, UT. Philadelphia (PA): AACR; Clin Cancer Res 2016;22(1_Suppl):Abstract nr 37.
Abstract 39: Association of single nucleotide polymorphisms (SNPs) in STS and SULT2B1 and response to androgen deprivation therapy (ADT) in men with advanced hormone sensitive prostate cancer (aHSPC)
Authors: Agarwal Neeraj , Buckley Tyler , Farnham James , Patel Shiven B. , Agarwal Archana , Tantravahi Srinivas , Teerlink Craig , Albright Frederick S. , Stephenson Robert A. , Alex Anitha , Cannon-Albright Lisa
Published: May 2016

Background: Germline variations in genes involved in androgen biosynthesis and metabolic pathways may predict time to response to androgen deprivation therapy (ADT) in aHSPC, serve as prognostic and predictive biomarkers, and guide towards more individualized upfront therapy.

Methods: 795 single nucleotide polymorphisms (SNPs) from the Illumina OmniExpress genotyping platform within the boundaries of 60 genes reported to be involved in the androgen metabolic pathway were investigated for association with time to onset of castration resistance prostate cancer (CRPC) in 118 Caucasian men with aHSPC, i.e. those progressing after definitive treatment, undergoing treatment with ADT. Cox proportional hazard analysis was employed using Gleason score as a covariate and assessing each SNP under an additive genetic model in which the number of minor alleles contributes increasing risk (or protection).

Results: Three SNPs in STS (steroid sulfatase gene), and three SNPs in SULT2B1 (sulfotransferase 2B1 gene) were significantly associated with time to CRPC on ADT while controlling for Gleason Score (p < .005).

Conclusions: SNPs in STS and SULT2B1 significantly associated with time to CRPC on ADT in aHSPC, and may serve as predictive markers to ADT in this setting. Data are being validated in a larger cohort.

Citation Format: Neeraj Agarwal, Tyler Buckley, James Farnham, Shiven B. Patel, Archana Agarwal, Srinivas Tantravahi, Craig Teerlink, Frederick S. Albright, Robert A. Stephenson, Anitha Alex, Lisa Cannon-Albright. Association of single nucleotide polymorphisms (SNPs) in STS and SULT2B1 and response to androgen deprivation therapy (ADT) in men with advanced hormone sensitive prostate cancer (aHSPC). [abstract]. In: Proceedings of the AACR Precision Medicine Series: Integrating Clinical Genomics and Cancer Therapy; Jun 13-16, 2015; Salt Lake City, UT. Philadelphia (PA): AACR; Clin Cancer Res 2016;22(1_Suppl):Abstract nr 39.
Abstract 42: Target inhibitory networks and drug response modeling
Authors: Shankavaram Uma , Camphausen Kevin
Published: May 2016

Cancer is a complex disease caused by multiple factors, which hamper effective drug discovery. Drug combination or multi target agents provide an alternate way to effectively modify disease networks. Synergistic drug pairs have special potential for treatment since they allow a desired effect to be achieved with lower total dose of administered medicine and usually with fewer side effects. However, the major challenge has been the prediction of chemotherapeutic efficacy based on the biological profile of the tumor.

Because of the lack of gene expression data treated with drug combinations, in this study, we present a computational approach to identify effective drug combinations by exploiting high throughput data. We constructed tissue specific network pharmacology based on large-scale screening data on drug treatment efficacies of 130 drugs under clinical and preclinical investigation and drug-target binding affinities. We evaluated CNS cancer subset and applied logic based network algorithm to predict effective drug combinations based on drug-target interactions and single drug sensitivity profiles.

Cancer cell based target inhibition network analysis in two case studies using glioma cell lines, (U87 and U251) identified distinct cell line survival pathways (p < 0.001), including cell proliferation, adhesion and growth factor signaling. We estimated pairwise drug synergy scores for all the target genes and identified several synergistic pairs with potential clinical relevance. Target inhibition modeling allowed systematic exploration of functional interactions between drugs and their targets to maximally inhibit multiple survival pathways.

Citation Format: Uma Shankavaram, Kevin Camphausen. Target inhibitory networks and drug response modeling. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Integrating Clinical Genomics and Cancer Therapy; Jun 13-16, 2015; Salt Lake City, UT. Philadelphia (PA): AACR; Clin Cancer Res 2016;22(1_Suppl):Abstract nr 42.
Abstract 47: Nucleophosmin/B23 (NPM/B23) and estrogen receptor alpha (ER alpha) in endometrial cancer
Authors: Lin Chiao Yun , Chao Angel , Wang Tzu-Hao , Lai Chyong-Huey
Published: May 2016

Purpose: Unopposed estrogen exposure is an important factor in the tumorigenesis of endometrial cancer. We aim to investigate the regulatory role of NPM/B23 in estrogen signaling in endometrial cancer.

Methods: To elucidate the underlying mechanism, we performed RT-qPCR, western blotting, promoter assay, immunoprecipitation, proliferation assay, protein stability assay, and ubiquitination assay in endometrial cancer cell lines.

Results: NPM/B23 was required for estrogen-induced endometrial proliferation, and the increase in NPM/B23 was estrogen receptor alpha (ER alpha)-dependent. Furthermore, estrogen increased NPM/B23 protein levels by repressing its ubiquitination and subsequently stabilizing the protein. The overexpression of alternate reading frame (ARF) suppressed the estrogen-induced increase in the NPM/B23 protein levels, indicating that ARF inhibited the observed estrogen-mediated NPM/B23 stabilization. Suppression of the NPM/B23-ARF interfered with the interaction and the subsequent increase in NPM/B23 protein levels.

Conclusions: Characterization of NPM/B23 in estrogen-mediated cell proliferation may extend our understanding of the tumorigenesis of endometrial cancers. Targeting B23 with hormonal therapy can be further explored for the treatment of endometrial cancer.

Citation Format: Chiao Yun Lin, Angel Chao, Tzu-Hao Wang, Chyong-Huey Lai. Nucleophosmin/B23 (NPM/B23) and estrogen receptor alpha (ER alpha) in endometrial cancer. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Integrating Clinical Genomics and Cancer Therapy; Jun 13-16, 2015; Salt Lake City, UT. Philadelphia (PA): AACR; Clin Cancer Res 2016;22(1_Suppl):Abstract nr 47.
Abstract 54: TOMA OS-seq: A high efficiency targeted re-sequencing strategy for detecting gene amplifications, rearrangements, indels and point mutations in tumor DNA isolated from FFPE solid tumors and plasma
Authors: Bouhlal Yosr , McKenzie Alexander , So Austin P.
Published: May 2016

Critical genetic errors, namely specific mutations, are required for the development and maintenance of cancer. Molecular diagnostics that leverage the breadth of next-generation sequencing (NGS) to identify this mutational landscape are therefore becoming an important clinical tool in the precision treatment of cancer. However, widespread adoption of NGS technologies in the clinic is hampered by the complexity of the NGS workflow, and the resulting limits in analytical capabilities arising from noise introduced by poor step-wise yields and bias from extensive application of PCR. To address these deficiencies, we introduce TOMA OS-Seq, a complete targeted resequencing workflow designed specifically for the requirements of the clinical laboratory. Based on oligo-selective sequencing (OS-seq), a panel of 96 genes - each with diagnostic or prognostic value - can be sequenced end-to-end from as little as 1 ng of input material, enabling the analysis of DNA purified from fine-needle aspirates and plasma. Utilizing a minimal number of high efficiency processing steps to create a targeted library, this simple workflow minimizes the requirement for PCR amplification, preserving the quantitative relationship among the loci interrogated in the original sample. TOMA OS-Seq is therefore capable of detecting copy number alterations, in addition to rearrangements/fusions, insertions/deletions, and single nucleotide variants.

Citation Format: Yosr Bouhlal, Alexander McKenzie, Austin P. So. TOMA OS-seq: A high efficiency targeted re-sequencing strategy for detecting gene amplifications, rearrangements, indels and point mutations in tumor DNA isolated from FFPE solid tumors and plasma. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Integrating Clinical Genomics and Cancer Therapy; Jun 13-16, 2015; Salt Lake City, UT. Philadelphia (PA): AACR; Clin Cancer Res 2016;22(1_Suppl):Abstract nr 54.
Abstract IA02: Precision medicine in acute lymphoblastic leukemia
Authors: Mullighan Charles G.
Published: May 2016

Acute lymphoblastic leukemia (ALL) is the commonest childhood tumor. Despite overall favorable treatment outcomes in children, relapsed ALL remains a leading cause of cancer death in young people. Current treatment regiments are mostly non-targeted and accompanied by significant toxicities, and new treatments tailored to patient-specific tumor vulnerabilities are required to further improve treatment outcomes. ALL has long been known to be genetically diverse. Recent genomic sequencing efforts have provided multiple new insights into the classification and genetic basis of ALL. ALL is now known to comprise multiple subtypes that have distinct constellations of chromosomal rearrangements, DNA copy number alterations and sequence mutations that perturb a limited number of key cellular pathways including lymphoid development, Ras and kinase signaling, tumor suppression drug metabolism and epigenetic regulation. These findings have several implications for precision medicine approaches.

First, different subtypes have distinct associations with relapse risk that will enable tailoring of the intensity of therapy, the requirement for intensive treatment approaches such as transplantation, and the potential for pathway-directed approaches such as BCL2 inhibitors, PI3K inhibitors and Ras inhibitors. Secondly, several subtypes have genetic alterations that are logical candidates for genomic testing and individualized targeted therapy. An important recent advance has been the identification of Ph-like ALL, a common high risk subtype of leukemia characterized by a range of genetic alterations activating cytokine receptor and kinase signaling amenable to ABL-class and JAK-STAT inhibitors. Large precision medicine trials identifying Ph-like ALL, the underlying genetic alterations, and directing patients to targeted kinase inhibitor therapy are currently in development. Thirdly, studies sequencing sequential samples obtained during therapy have identified several genes and pathways selectively mutated at relapse, including epigenetic regulators, that are logical targets for therapeutic intervention. Finally, genome sequencing is being used to develop highly sensitive approaches to detect and track response to treatment. Examples of each of these paradigms will be presented that highlight the potential of precision medicine to further improve treatment outcomes in ALL.

Citation Format: Charles G. Mullighan. Precision medicine in acute lymphoblastic leukemia. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Integrating Clinical Genomics and Cancer Therapy; Jun 13-16, 2015; Salt Lake City, UT. Philadelphia (PA): AACR; Clin Cancer Res 2016;22(1_Suppl):Abstract nr IA02.
Abstract IA04: Functional genomics for discovery of pathogenic mechanisms of disease and novel therapies for hematologic malignancies
Authors: Tyner Jeffrey W.
Published: May 2016

Our ability to quickly and cost-effectively decode cancer genomes and identify somatic mutations has been revolutionized by deep sequencing technologies. In some instances, the identification of these somatic driver mutations has resulted in implementation of more effective therapeutic strategies for patients. However, in many instances knowledge of somatic drivers has not resulted in new therapeutic strategies. In some cases, these driver mutations are gain-of-function and targeted drugs have not yet been developed, such that further drug development will bridge the translational gap between genomic knowledge and clinical translation. However, in many cases the somatic driver mutations are not directly targetable because the mutations result in loss-of-function of the mutated gene. In these cases, we face a critical knowledge gap between understanding of the identity of molecular drivers of disease and understanding of the dysregulated signaling pathways that result from these loss-of-function genetic events. It will be important to close this knowledge gap in order to better understand the targeted therapies that can be effective for all somatic driver events, such that all cancer patients can receive effective, individually tailored therapies. My lab has developed ex vivo, functional screening platforms that can be applied directly to primary specimens from patients with hematologic malignancies. These assays are based on panels of drugs and can deliver information about the specific drugs (or combinations of drugs) to which a patient's tumor cells are hypersensitive. Integration of these functional data with genomic information about each patient's tumor cells has allowed us to accelerate the process by which we discover actionable therapeutic strategies that correlate with tumor genotypes. In addition, these assays are completed in a clinically relevant time frame of three days, such that the data from these assays can be used to directly inform therapeutic strategies, even in the absence of genomic information. In this lecture, I will provide an overview of these functional screening platforms, I will illustrate strategies of data integration that have enabled rapid discovery and translation of new targeted therapy strategies, and I will discuss our experience to date with direct clinical implementation of findings from these functional screening data.

Citation Format: Jeffrey W. Tyner. Functional genomics for discovery of pathogenic mechanisms of disease and novel therapies for hematologic malignancies. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Integrating Clinical Genomics and Cancer Therapy; Jun 13-16, 2015; Salt Lake City, UT. Philadelphia (PA): AACR; Clin Cancer Res 2016;22(1_Suppl):Abstract nr IA04.
Abstract IA05: Clinical genomics alters care, informs new science
Authors: Taylor Barry S.
Published: May 2016

Broad-based prospective sequencing has begun to inform the care of individual cancer patients. Such genomically driven cancer medicine is facilitating the use of mechanism-based therapeutic approaches in patients. In addition to the value of such data to inform the treatment of individual patients under active clinical care, over time these accrue as unprecedented datasets of integrated genomic, clinical, and treatment response data that can inform translational cancer research. Here, we present preliminary results of our efforts to combine both large-scale retrospective and prospective sequencing data in cancer to drive translational cancer biology. We will report on our efforts to develop a computational oncology framework to mine these data not only for new treatment hypotheses that can be tested clinically and the preliminary results from those studies, but also how this is driving new approaches to understand so-called long tail mutations both biologically and clinically. These efforts lie at the interface of retrospective and prospective studies to inform precision oncology through new tools that move beyond enumerating individual genetic vulnerabilities toward interpreting their biological and clinical interpretation for the benefit of patients.

Citation Format: Barry S. Taylor. Clinical genomics alters care, informs new science. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Integrating Clinical Genomics and Cancer Therapy; Jun 13-16, 2015; Salt Lake City, UT. Philadelphia (PA): AACR; Clin Cancer Res 2016;22(1_Suppl):Abstract nr IA05.
Abstract IA17: Delivering on the promise of personalized medicine
Authors: Mills Gordon B.
Published: May 2016

The realization of the promise of personalized molecular medicine requires efficient development and implementation of novel targeted therapeutics linked to molecular markers able to identify patients most likely to benefit. An efficient integration of DNA, RNA and protein information content will be needed to identify patients likely to respond to particular therapies. The plethora of aberrations present in each tumor, the need to distinguish drivers from passengers, and the combinatorial effects of aberrations on critical cellular functions represent key challenges. The overall likelihood of response to therapy represents the interaction between predictors of sensitivity and predictors of resistance. Resistance can be pre-existing, adaptive, or acquired. Resistance can also occur through heterogeneity of molecular changes within the tumor and metastases. Adaptive resistance, which is the consequence of activation of homeostatic loops and phylogenetically conserved stress responses, provides a potential therapeutic liability that can be leveraged for rational combinatorial therapy. Thus a comprehensive analysis of patient tumors before, during and after treatment should become the standard of practice. Testing these precepts will require the development and implementation of novel trial designs. It is likely that we will need to increase the size of phase I and II trials to allow the identification and validation of molecular markers at the same time as the initial evaluation the toxicity and efficacy of targeted therapeutics. This will come with the advantage much smaller phase III trials of patients selected for the likelihood to respond accelerating the approval of effective targeted therapeutics.

Citation Format: Gordon B. Mills. Delivering on the promise of personalized medicine. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Integrating Clinical Genomics and Cancer Therapy; Jun 13-16, 2015; Salt Lake City, UT. Philadelphia (PA): AACR; Clin Cancer Res 2016;22(1_Suppl):Abstract nr IA17.
Abstract A023: The clinical utility of the modified Glasgow Prognostic Score in patients undergoing potentially curative colorectal cancer resection
Authors: Park James H. , Roxburgh Campbell SD , Horgan Paul G. , McMillan Donald C.
Published: May 2016

The aim of the present study was to examine the clinical utility of assessment of the systemic inflammatory response, using the modified Glasgow Prognostic Score (mGPS), in combination with TNM stage in 1000 consecutive patients undergoing potentially curative colorectal cancer resection in a single institution. An mGPS of 0 (CRP<10mg/L), 1 (CRP>10mg/L/ albumin<35g/L) and 2 (CRP>10mg/L/ albumin<35g/L) was observed in 63%, 21% and 16% of patients respectively. The mGPS was associated with reduced cancer-specific survival (P<0.01), independent of TNM stage; the combination of TNM stage and mGPS stratified five-year survival from 97% (TNM I, mGPS=0) to 32% (TNM III, mGPS=2) (P<0.001). Furthermore, in patients undergoing elective resection of TNM III colon cancer, mGPS stratified survival of patients who received adjuvant chemotherapy from 91% (mGPS=0) to 56% (mGPS=2) (P=0.001). Indeed, the mGPS is routinely available and, in combination with TNM-based staging, provides additional prognostic information following potentially curative colorectal cancer resection.

Citation Format: James H. Park, Campbell SD Roxburgh, Paul G. Horgan, Donald C. McMillan. The clinical utility of the modified Glasgow Prognostic Score in patients undergoing potentially curative colorectal cancer resection. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr A023.
Abstract A027: A phase I open-label, non-randomized study of recombinant compound interferon (rSIFN-co) in patients with advanced solid tumors
Authors: Choo Su Pin , Tai Wai Meng , Macapagal Maria , Ahmad Aziah , Li Greg , Goh Mui Leng , Ng Matthew , Tan Tira , Tan Shao Weng , Lim Wan Teck
Published: May 2016

Background: Recombinant compound interferon (rSIFN-co) is a new class of interferon derived from altering the spatial structure and configuration of the basic interferon protein. rSIFN-co displayed greater anti-tumor activity in solid tumors compared to interferon α-2b in pre-clinical studies.

Methods: Patients with advanced malignancies were enrolled to determine the safety, tolerability, recommended phase II dose (RP2D), pharmacodynamics (PD) and preliminary anti-tumor activities in a 3+3 dose escalation design. Two pre-defined dose levels were planned. rSIFN-co was dosed 3 times a week via subcutaneous injection for 21 days followed by 7 days rest. This was preceded by a lead in phase. PD studies comprised of FDG-PET, pre- and post treatment changes in plasma cytokine profile and tumor repressive/ enhancing genes as well as 2'5'-oligoadenylate synthetase levels.

Results: Results presented are based on preliminary data collected as of Jan. 27, 2015. 22 patients (median age 65.5 years, 14 male and 8 female) have been enrolled with 21 evaluable for safety and 15 amenable to efficacy analysis. 10 hepatocellular carcinoma (HCC), 6 non-small cell lung cancer (NSCLC), 4 colorectal cancer (CRC), 1 melanoma and 1 nasopharyngeal carcinoma patients (pts) were dosed at 2 dose levels. The RP2D was 24μg 3x/week for 21days followed by 7 days rest. No dose limiting toxicities were encountered. Most adverse events (92%) were CTCAE grade 1 and 2. The most frequent grade 1/2 treatment related adverse events (trAEs) were flu-like symptoms (90%), anorexia (33%), fatigue (33%), weight loss (33%), nausea (29%), thrombocytopenia (29%), and diarrhea (24%). Grade 3/4 trAEs were fever (2 pts), leucopenia (2 pts), transaminitis (2 pts), weight loss (1 pt), fatigue (1 pt), afebrile neutropenia (1 pt) and syncope (1 pt). 11 patients had stable disease (SD) and 4 patients had progressive disease (PD). 4 patients (2 HCC, 1 NSCLC and 1 CRC) had prolonged stable disease (>16 weeks). Detailed PD data will be presented for all cohorts.

Conclusion: The RP2D was 24μg three times a week for 21days followed by 7 days rest with a lead in phase. Modest activity was seen in heavily pre-treated advanced solid tumor patients.

Citation Format: Su Pin Choo, Wai Meng Tai, Maria Macapagal, Aziah Ahmad, Greg Li, Mui Leng Goh, Matthew Ng, Tira Tan, Shao Weng Tan, Wan Teck Lim. A phase I open-label, non-randomized study of recombinant compound interferon (rSIFN-co) in patients with advanced solid tumors. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr A027.
Abstract A058: GITRL-Fc, an immunotherapeutic agent that stimulates T-cell-mediated antitumor immune response
Authors: Axelrod Fumiko , Jie Hyun-Bae , Mayes Erin , Monteon Jorge , Srivastava Minu , Yun Rui , Park Inkyung Angie , Gurney Austin
Published: May 2016

GITRL, (Glucocorticoid-Induced Tumor Necrosis Factor Receptor Ligand, TNFSF18) is a member of the TNF family of ligands and naturally exists as a membrane-anchored type II protein that self assembles as a trimer. GITRL activates the co-stimulatory receptor GITR. A novel single-gene linkerless GITRL trimer was shown to be functional when fused to either the N- or C-terminus of an immunoglobulin Fc domain, offering a flexible strategy that may also be amenable to the production of bispecific agents. GITRL-Fc activated GITR signaling more effectively than prototype GITR agonist antibody DTA-1. GITRL-Fc promoted a robust anti-tumor immune response in several murine tumor graft models, including the apparent total regression of some treated tumors. GITRL-Fc potentiated tumor specific T-cell responses, particularly of the Th1 type, increased antigen-specific CD8 response, and promoted a reduction in Treg-mediated immune-suppressive activity.

Citation Format: Fumiko Axelrod, Hyun-Bae Jie, Erin Mayes, Jorge Monteon, Minu Srivastava, Rui Yun, Inkyung Angie Park, Austin Gurney. GITRL-Fc, an immunotherapeutic agent that stimulates T-cell-mediated antitumor immune response. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr A058.
Abstract A067: Quality of initial activation of CD8 T cells specific for TRP1 determines long-term antitumor immunity
Authors: Tyler Paul , Servos Mariah , Dougan Stephanie K.
Published: May 2016

We used somatic cell nuclear transfer to generate two independent lines of transnuclear mice from CD8 T cells recognizing the endogenous melanoma antigen TRP1. These two lines, while having the same specificity, differ by 100 fold in their affinity for antigen. We previously reported that activated CD8 T cells from either TRP1high or TRP1low mouse lines have anti-tumor activity in vitro and can slow the progression of B16 melanoma in vivo to similar extents, suggesting that TCR affinity is not necessarily correlated with anti-tumor function.

Here we first analyzed the mechanism of tumor control in our TRP1 model. In tumor-bearing mice receiving activated TRP1 cells by adoptive transfer, we examined tumor infiltrates and found 1) an increase in the CD8:Foxp3 ratio; 2) a decrease in myeloid suppressor cells; and 3) greatly increased expression of MHC class I and class II expression by the B16 tumor cells. B16 cells upregulate MHC expression in vitro in response to IFNγ; thus, we hypothesize that MHC expression on B16 cells in mice receiving TRP1 adoptive cell therapy is a signature of IFNγ production in the tumor.

We further used a panel of altered peptide ligands to extend the range of TCR activation potentials in our TRP1 model. One ligand, K8, binds poorly to MHC class I yet stimulates robust proliferation, cytotoxicity and cytokine production from both TRP1high and TRP1low cells. Interestingly, K8-activated TRP1high and TRP1low cells fail to upregulate PD-1 expression. When adoptively transferred into tumor-bearing mice, K8-activated TRP1 cells delayed tumor growth to a similar extent as TRP1 cells activated by the reference peptide, yet mice receiving K8-activated cells had an increased chance of developing long-term memory. We show TRP1 tetramer+ CD44+ CD8+ T cells in K8-treated mice that survived both primary and secondary challenge with B16. Transcriptional profiling of K8 versus reference peptide activated TRP1 cells reveals a transcriptional program associated with anti-tumor immunity.

Citation Format: Paul Tyler, Mariah Servos, Stephanie K. Dougan. Quality of initial activation of CD8 T cells specific for TRP1 determines long-term antitumor immunity. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr A067.
Abstract A068: Imbalance in regulatory T (Treg) cells frequency in peripheral circulation of patients with oral squamous cell carcinoma and their effects on growth of oral cancer
Authors: Aggarwal Sadhna , Das Satya N. , Sharma Suresh C.
Published: May 2016

Regulatory T (Treg) cells play a major role in establishment and maintenance of self tolerance of immune system. Deficit in the numbers of Tregs contributes to the development of autoimmune diseases, allergy and graft rejection whereas abundance of Tregs suppresses anti-tumour immune response leading to tumour immuno-evasion. In this study, we have estimated the frequency of distinct immune cells, Tregs and its phenotypic subsets in patients with Oral Squamous Cell Carcinoma (OSCC) by multicolour flowcytometry. Subsequently we investigated the effects of MACS sorted Tregs on growth/proliferation/angiogenesis of OSCC cell lines in vitro.

Frequency of all the effector type anti-tumour immune cells such as CD8+ T-cells, NK (CD56+) and NKT (CD3+CD56+) cells was found to be decreased whereas CD3+, CD4+, γδ-T cells were upregulated in OSCC patients. We observed significantly increased (p < 0.05) prevalence of Tregs and their phenotypic subsets (CD4+CD25+, CD4+FoxP3+, CD8+FoxP3+, CD4+CD25+FoxP3+) in peripheral circulation of OSCC patients as compared to that of the normal individuals. Our study further showed a definite association between the OSCC progression and elevated Tregs frequency. An association between reciprocally regulated Treg and Th17 cells was also observed. The increased frequency of CD4+CD8+CD25+FoxP3+, a unique T cells subset was seen in OSCC patients. The CTLA-4+ and GITR+ Tregs also showed higher occurrence in patients indicating their contribution to the immuno-suppressive function of Tregs in OSCC. Introduction of MACS isolated Treg cells to in vitro oral cancer cell culture resulted in increased proliferation and expression of angiogenic marker VEGF by cancer cells showing that these suppressor cells helps in growth and proliferation of OSCC cells.

Accumulation of heterogeneous subsets of Tregs in the peripheral circulation indicates their suggestive role in tumour induced immuno-suppression as well as in growth and proliferation of oral cancer cells. Therefore, inhibitory manipulation of Tregs, in terms of their frequency and functional activity, can be added to the therapeutic armamentarium for enhancing anti-tumour immunity in OSCC patients and preventing the establishment and progression of oral cancer.

Citation Format: Sadhna Aggarwal, Satya N. Das, Suresh C. Sharma. Imbalance in regulatory T (Treg) cells frequency in peripheral circulation of patients with oral squamous cell carcinoma and their effects on growth of oral cancer. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr A068.
Abstract A078: Computational discovery and experimental validation of novel drug targets in immuno-oncology
Authors: Machlenkin Arthur , Levy Ofer , Rotman Galit
Published: May 2016

The past few years have witnessed a renaissance in the field of immuno-oncology largely due to the clinical success in targeting the immune checkpoints CTLA-4 and PD-1. Towards identification of novel immune checkpoint drug targets we developed a dedicated predictive discovery platform.

The B7/CD28 discovery platform is a predictive model based on genomic and protein features along with expression patterns of known B7/CD28 proteins. The platform has been tested and validated extensively and has demonstrated its validity by identifying non-novel immune checkpoints such as TIGIT and VISTA, which were not used in the design stage.

The B7/CD28 predictive platform was employed to identify 11 new immune checkpoint candidates which are currently in different validation stages. In this poster, we will describe our discovery approach as well as our validation path. In addition, we will present experimental data demonstrating the immuno-modulatory function and expression patterns of several of our novel immune checkpoints. These experimental results serve as an additional confirmation to the accuracy of our B7/CD28 predictive discovery platform and shed light on the therapeutic potential of the novel immune checkpoints identified using this unique discovery approach.

Citation Format: Arthur Machlenkin, Ofer Levy, Galit Rotman. Computational discovery and experimental validation of novel drug targets in immuno-oncology. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr A078.
Abstract A087: Phase separation of signaling molecules promotes T cell receptor signal transduction
Authors: Su Xiaolei , Ditlev Jon , Hui Enfu , Banjade Sudeep , Okrut Julia , Taunton Jack , Rosen Mike , Vale Ron
Published: May 2016

Signaling through the T cell receptor (TCR) mediates T cell activation, differentiation, and proliferation. Previous microscopy studies have shown the spatial reorganization of signaling molecules into microclusters on the plasma membrane after TCR activation. However, how microclusters form and what roles these structures play in signal transduction remain unclear. Importantly, direct comparison of the biochemical activities of signaling molecules in the clustered versus unclustered state has been lacking. Here, we address these questions by reconstituting a TCR signaling pathway in vitro with 12 purified proteins, beginning with TCR phosphorylation and culminating in actin polymerization. We show that upon initiation of phosphorylation, multivalent protein-protein interactions drive the phase separation of signaling molecules into microclusters that display liquid-like properties. These clusters enrich kinases but exclude phosphatases, thus sustaining protein phosphorylation. We also directly demonstrate that the reorganization of actin regulatory proteins into these clusters substantially enhances actin filament assembly at the membrane. These results establish that microclusters create unique physical and biochemical environments that promote reactions in TCR signaling. The principles revealed here likely apply to other systems that exploit phase separation to regulate biochemical reactions on membranes, in the cytoplasm, or the nucleus.

Citation Format: Xiaolei Su, Jon Ditlev, Enfu Hui, Sudeep Banjade, Julia Okrut, Jack Taunton, Mike Rosen, Ron Vale. Phase separation of signaling molecules promotes T cell receptor signal transduction. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr A087.
Abstract A105: Cross-talk between tumors can affect responses to therapy
Authors: Devaud Christel , Westwood Jennifer A. , John Liza , Yong Carmen S.M. , Beavis Paul A. , Snyder Linda A. , Schwendener Reto A. , Phillip Darcy K. , Kershaw Michael H.
Published: May 2016

Tumors are associated with a microenvironment that can influence responses to immunotherapy. Advanced stages of cancer often involve multiple tumors in different locations in the body. We have demonstrated that the tumor microenvironment can vary with anatomical site, and that normal tissue surrounding the site of tumor implantation can contribute to sculpting the subsequent tumor microenvironment. Furthermore, we demonstrate that tumor responses to immunotherapy, consisting of three agonist antibodies, vary with anatomical site. We also investigated whether tumors and their disparate microenvironment can interact with each other at a distance, in a multiple tumor setting, through a form of cross-talk, to affect their responses to immunotherapy. Our study investigated the cross-talk between two tumors with disparate microenvironments in a mouse model. We demonstrated that immunosuppressive visceral tumors could influence distant subcutaneous (SC) tumors, normally responsive to immunotherapy, to render them resistant. We observed distinct modifications in the SC tumor microenvironment following cross-talk with kidney tumors, which exhibit a type-2 macrophage-related immunosuppressive microenvironment. Indeed, when a concomitant kidney tumor was present in the mouse, the SC tumors were highly infiltrated with M2 macrophages and had a reduced T cell and NK cell effector immune profile. Finally, blocking the M2-associated chemokine CCL2 or depleting macrophages, significantly improved the effect of immunotherapy on SC tumors in the presence of concomitant kidney tumors. This work presents a novel finding describing the potential negative influence that a tumor, with a strong immunosuppressive microenvironment, can exert on distant tumors that would normally be responsive to treatment. This report may lead to a new outlook in the selection and prioritization of treatment of multiple tumors in advanced metastatic cancer, which will consider the anatomical location and corresponding microenvironment of individual tumors.

Citation Format: Christel Devaud, Jennifer A. Westwood, Liza John, Carmen S.M. Yong, Paul A. Beavis, Linda A. Snyder, Reto A. Schwendener, Darcy K. Phillip, Michael H. Kershaw. Cross-talk between tumors can affect responses to therapy. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr A105.
Abstract A110: Mutant MHC class II epitopes drive therapeutic immune responses to cancer
Authors: Vormehr Mathias , Kreiter Sebastian , Roemer Niels van de , Diken Mustafa , Löwer Martin , Vascotto Fulvia , Diekmann Jan , Boegel Sebastian , Schroers Barbara , Tadmor Arbel D. , Türeci Özlem , Sahin Ugur
Published: May 2016

Mutations are regarded as ideal targets for cancer immunotherapy. As neoepitopes with strict lack of expression in any healthy tissue, they are expected to be safe and could bypass the central tolerance mechanisms. Recent advances in nucleic acid sequencing technologies have revolutionized the field of genomics, allowing the readily targeting of mutated neoantigens for personalized cancer vaccination.

We demonstrated in three independent murine tumor models that a considerable fraction of non-synonymous cancer mutations is immunogenic and that unexpectedly the immunogenic mutanome is pre-dominantly recognized by CD4+ T cells. RNA vaccination with such MHC class II restricted immunogenic mutations leads to infiltration of CD4+ and CD8+ T cells into the tumor, reduces intratumoral regulatory T cells and ultimately confers strong anti-tumor activity. Encouraged by these findings we set up a process comprising mutation detection by exome sequencing, selection of vaccine targets by solely bioinformatical prioritization of mutated epitopes predicted to be abundantly expressed and presented on MHC class II molecules. Synthetic mRNA vaccines encoding multiple of these prioritized mutated epitopes induce potent tumor control and complete rejection of established aggressively growing tumors in mice. Moreover, we demonstrate that CD4+ T cell neoepitope vaccination primes CTL responses against an independent immunodominant antigen in tumor bearing mice indicating orchestration of antigen spread. Our findings reveal that cancer mutation based MHC class II restricted epitopes are attractive vaccination targets and provide the preclinical proof of concept for an integrated process from tumor sample to a cancer vaccine customized to the unique repertoire of each patient`s tumor.

Citation Format: Mathias Vormehr, Sebastian Kreiter, Niels van de Roemer, Mustafa Diken, Martin Löwer, Fulvia Vascotto, Jan Diekmann, Sebastian Boegel, Barbara Schroers, Arbel D. Tadmor, Özlem Türeci, Ugur Sahin. Mutant MHC class II epitopes drive therapeutic immune responses to cancer. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr A110.
Abstract A118: Transforming growth factor-beta as a therapeutic target for pancreatic cancer
Authors: Meraz Ismail , Vergara Leoncio , Suki Sarah , Gunamalai Prem Siddharth , Serda Rita Elena
Published: May 2016

Recent evidence supports a strong role for immune suppression in facilitating cancer progression. Transforming growth factor beta (TGFß) regulates a diverse array of biological events, including neoangiogenesis, stromal formation, and immune responses. Both TGFß and a small molecule inhibitor of TGFßR-1, LY364947, were effective regulators of these processes in vitro. Using quantitative RT-PCR and fluorescence imaging, TGFß increased collagen production by pancreatic stellate cells while addition of LY364947 suppressed production below basal levels. Endothelial cell proliferation, measured as a function of endothelial tube formation in Geltrex® matrix, was similarly increased by TGFß and reduced to basal levels by LY364947. To explore the in vivo effects of these molecules, LY364947 was encapsulated in liposomes to encourage tumor accumulation and to avoid administration of DMSO vehicle. Liposomes were prepared with a molar ratio of 6:3:1 1,2-dipalmitoyl-sn-glycero-3-phosphocholine /Chol/1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] using a Hei-Vap Series Heidolph Rotatory Evaporator and a Thermobarrel Extruder. In contrast to in vitro studies, tumor stromal content in human pancreatic capan-2 tumors established in nude mice was not influenced by LY364947, free or liposomal. In addition, tumor progression, as a function of size, and myeloid populations were unaltered by weekly treatments with liposomal LY364947. Thus the influence of TGFßR-1 inhibition in cell cultures does not translate directly into biologically equivalent events.

Citation Format: Ismail Meraz, Leoncio Vergara, Sarah Suki, Prem Siddharth Gunamalai, Rita Elena Serda. Transforming growth factor-beta as a therapeutic target for pancreatic cancer. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr A118.
Abstract A120: Tumor-educated B cells acquire LAP/TGF-β1 and PD-L1 expression and suppress antitumor immune response
Authors: Zhang Yu , Morgan Richard , Chen Chuan , Cai Yancheng , Shin Seung-Uon , Cho Hyun-Mi , Bayati Ahmed Al , Pimental Augustin , Rosenblatt Joseph D.
Published: May 2016

B lymphocytes play a role in inhibiting the immune response against certain tumors, but the underlying mechanisms are poorly understood. EMT-6 mammary tumors grow well in wild type (WT) mice but show reduced growth in B-cell deficient μ-/- BALB/c mice (BCDM). WT mice demonstrate B cell infiltration into the tumor bed and a reduced cytolytic T cell response relative to BCDM. Expression of LAP/TGF-β1, CD80, CD86 and PDL-1 is increased in tumor infiltrating B cells (TIL-B) relative to splenic B cells. LAP/TGF-β1 expression progressively increased from 5.4±1.7% on d8 to 43.1±6.1% of TIL-B by d21 post tumor implantation. Co-culture of EMT-6 tumor cells with naïve B cells generated B cells (EMT6-B) with a similar immunophenotype to TIL-B. Purified TIL-B or in vitro generated EMT6-B markedly suppressed CD4+CD25- T cell proliferation, and Th1 cytokine secretion compared to naïve splenic B cells. Acquired B regulatory function required direct tumor cell: B cell contact, and was partially reversed by antibody to TGF-β1 or PDL-1, leading to tumor rejection in vivo. B cell acquisition of a suppressive phenotype following tumor infiltration may result in profound inhibition of T cell anti-tumor response. Similar B cell mediated immunosuppression may be operative in human solid tumors.

Citation Format: Yu Zhang, Richard Morgan, Chuan Chen, Yancheng Cai, Seung-Uon Shin, Hyun-Mi Cho, Ahmed Al Bayati, Augustin Pimental, Joseph D. Rosenblatt. Tumor-educated B cells acquire LAP/TGF-β1 and PD-L1 expression and suppress antitumor immune response. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr A120.