bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5495: BET bromodomain inhibitors abrogate cell cycle progression and induces apoptosis in Myc-induced mouse lymphoma cells without affecting MYC transcription
Authors: Bhadury Joydeep , Nilsson Lisa M. , Green Lydia , Li Zhoulei , Gesner Emily M. , Muralidharan Somsundar Veppil , Hansen Henrik C. , Keller Ulrich B. , Mclure Kevin G. , Nilsson Jonas A.
Published: May 2015

Overexpression of one of the three MYC genes is a hallmark of many human cancers. They encode transcription factors that regulate expression of a number of genes critical for tumor development. Conditional transgenic mouse models have shown that Myc inhibition causes tumor regression; Myc is therefore an attractive target for therapy, and effective pharmacological Myc inhibition has been a long-standing goal in cancer research. Recent publication have shown that selective inhibition of BET bromodomain and extra-terminal (BET) domain family of proteins, that bind to acetylated lysines on histones, show promise as potent anti-cancer drugs via down-regulation of the MYC oncogene. Here we confirm that two structurally different BET protein inhibitors (BETi), JQ1 (proto-type) and RVX2135, inhibit the proliferation and induce apoptosis of lymphoma cells arising in Myc-transgenic mice. BETi have been reported to directly inhibit MYC transcription. Surprisingly, in our system BET inhibition had no effect on MYC transcription, despite exhibiting broad transcriptional effects evident from expression profiling. Our data challenge the prevailing view that BETi operate primarily via suppression of MYC transcription in hematological malignancies. Instead we suggest that BET proteins have Myc-independent pleiotropic effects that should be exploited to treat a wide range of cancers. Myc still remains a challenging target for therapy to date.

Citation Format: Joydeep Bhadury, Lisa M. Nilsson, Lydia Green, Zhoulei Li, Emily M. Gesner, Somsundar Veppil Muralidharan, Henrik C. Hansen, Ulrich B. Keller, Kevin G. Mclure, Jonas A. Nilsson. BET bromodomain inhibitors abrogate cell cycle progression and induces apoptosis in Myc-induced mouse lymphoma cells without affecting MYC transcription. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5495. doi:10.1158/1538-7445.AM2014-5495
Abstract CT102: Efficacy and toxicity management of 19-28z CAR T cell therapy in B cell acute lymphoblastic leukemia
Authors: Sadelain Michel , Brentjens Renier , Davila Marco , Riviere Isabelle , Wang Xiuyan , Bartido Shirley , Park Jae , Bouhassira Diana , Curran Kevin , Chung Stephen , Stefanski Jolanta , Borquez-Ojeda Oriana , Giralt Sergio
Published: May 2015

Chimeric antigen receptors (CARs) are artificial receptors for antigen that redirect antigen specificity, activate T cells and further enhance T cell function through their costimulatory component. Three groups, including our own, have reported objective tumor responses when infusing autologous T cells genetically modified with CD19-targeted CARs into patients with chronic lymphocytic leukemia (CLL), other indolent non-Hodgkin lymphomas (NHL) and, most dramatically in patients with relapsed or refractory B cell acute lymphoblastic leukemia (B-ALL). Here we report on 16 patients with relapsed or refractory B-ALL that we treated with autologous T cells expressing the 19-28z CAR specific to the CD19 antigen. The overall complete response rate was 88%, as assessed by morphological criteria and IgH deep sequencing. This allowed us to transition a majority of these patients to a standard of care allogeneic hematopoietic stem cell transplant (allo-SCT). This therapy was as effective in high-risk patients with Ph+ disease as in those with relapsed disease following prior allo-SCT. Through systematic analysis of clinical data and serum cytokine levels over the first 21 days following T cell infusion, we have defined diagnostic criteria for a severe cytokine release syndrome (sCRS), with the goal of better identifying the subset of patients who will likely require therapeutic intervention with corticosteroids or interleukin-6 receptor blockade to curb the CRS. Additionally, we found that serum C-reactive protein (CRP), a readily available laboratory study, can serve as a reliable indicator for the severity of the CRS and a road map for patient management at centers now contemplating the use of CAR T cell therapy. Based on these remarkably robust clinical results and the toxicity management algorithm we present here, we will soon open a multi-center Phase II clinical trial to further evaluate the efficacy of 19-28z CAR T cells and prospectively validate our proposed CRS monitoring and intervention guidelines in patients who are treated with CAR therapy.

Citation Format: Michel Sadelain, Renier Brentjens, Marco Davila, Isabelle Riviere, Xiuyan Wang, Shirley Bartido, Jae Park, Diana Bouhassira, Kevin Curran, Stephen Chung, Jolanta Stefanski, Oriana Borquez-Ojeda, Sergio Giralt. Efficacy and toxicity management of 19-28z CAR T cell therapy in B cell acute lymphoblastic leukemia. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr CT102. doi:10.1158/1538-7445.AM2014-CT102
Abstract CT228: A phase II study of afatinib (A) in patients (pts) with metastatic human epidermal growth factor receptor (HER2)-positive trastuzumab (T) refractory esophagogastric (EG) cancer
Authors: Janjigian Yelena Y. , Imtiaz Tooba , Ilson David H. , Solit David B. , Berger Michael F. , Vakiani Efsevia
Published: May 2015

Background: Trastuzumab (T) combined with chemotherapy has been the standard of care for pts with HER2+ EG cancer. Resistance to T is now emerging in this population. Afatinib (A), a potent ErbB Family Blocker, induced nearly complete tumor regression in MSKCC HER2+ patient derived xenografts (PDX). We report the initial results of a phase II study of afatinib in patients with T refractory EG cancer.

Methods: Pts with HER2+ (IHC 3+ or FISH>2.0) EG cancer -progressive on trastuzumab -received A 40 mg. Archival pre-T tissue, tumor biopsy after progression on T and after 1 week on A mandated on protocol. The primary endpoint-overall disease control: stable disease (SD) or partial response (PR).

Results: 14 pts treated with A; median duration 5.1 mos (1.7 to 12.1 mos), median 2 (1 to 4) prior T regimens, 64% of tumors IHC3+; 36% IHC2+/FISH>2.2. Adverse events included: diarrhea (Grade 1/2:69%), fatigue (Grade1/2: 54%), rash (Grade 1/2:54%), mucositis (Grade1: 23%), paronychia (Grade 1/2:15%). 4 of 14 pts (28%) with disease control (PR or SD); including 1 pt with confirmed PR - a durable 75% regression of biopsy proven metastases. Median OS 6.6 mos (1.9 to NR). PDXs established from biopsies of T refractory tumors of 5 pts. EGFR amplification was detected in the tumor of the patient with PR on afatinib, suggesting EGFR amplification as a predictor of response to afatinib. High level of HER2 amplification was retained in post-trastuzumab resistance sample suggesting ongoing dependence on HER2. PIK3CA mutations were observed in tumors of 33% (2 of 6) patients, ERBB3 mutations in 33% cases (2 of 6), and MTOR or FBXW7 mutations in 50% (3 of 6) patients.

Conclusions:

Afatinib shows clinical efficacy in patients with T refractory EG cancer. The study met the criteria for expansion of the accrual based on original Simon's minimax two-stage design. Efforts to elucidate the mechanisms of T resistance including validation of potential drivers of T resistance using HER2+ PDXs are ongoing. Updated molecular and clinical data will be presented.

Citation Format: Yelena Y. Janjigian, Tooba Imtiaz, David H. Ilson, David B, Solit, Michael F. Berger, Efsevia Vakiani. A phase II study of afatinib (A) in patients (pts) with metastatic human epidermal growth factor receptor (HER2)-positive trastuzumab (T) refractory esophagogastric (EG) cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr CT228. doi:10.1158/1538-7445.AM2014-CT228
Abstract CT411: Immune profiling of patients vaccinated with the survivin targeted therapeutic vaccine DPX-Survivac demonstrates durable polyfunctional CD4+ and CD8+ T cells in ovarian cancer patients
Authors: Peretz Yoav , Sauvé Dominike , Gagnon Dominic , Khan Salim Ahmed , Lévesque Geneviève , Saha Nathalie , Croteau Gilbert , Hébert Valérie , Savard Karyne , Diab Bader Yassine , Poulin Jean-Francois , Landry Claire , Karkada Mohan , Mansour Marc
Published: May 2015

Survivin is a member of the inhibitor of apoptosis protein family and its over-expression has been reported in many cancer types such as melanoma, colon, brain, breast, and ovarian cancers making it a promising target for immunotherapy. DPX-Survivac is a Survivin multi-epitope vaccine that covers a broad range of HLA haplotypes (HLA-A1, A2, A3, A24, B7). The adjuvanted water free depot vaccine is designed to specifically induce CD8+ T cells. In a Phase I study in ovarian cancer patients (n=18), we aimed to evaluate one dose level of the vaccine (0.5 mL) without cyclophosphamide (Cohort A) and two dose levels (0.1 mL and 0.5 mL) combined with oral low dose cyclophosphamide as an immune modulator (Cohorts B and C). Using multiparametric flow cytometry, we measured the ex-vivo frequency, phenotype and function of Regulatory T cells (CD25+FoxP3+Ki67+) and Survivin-specific CD4 and CD8 T cells. Our data shows that cohort C preferentially mounted survivin-specific CD8 T cells which significantly peaked at one month following the third vaccination (study day 70) and persisted until at least day 126 post-vaccination. This polyfunctional T cell response was endowed with cytotoxic and proliferative functions as measured by the expression of IL-2, IFNγ, TNFα, Granzyme B and CD107a. The immune responses identified in this cohort functionally diversified over time and were distributed within the Effector memory (CD45RA-CD27-), Central Memory (CD45RA-CD27+) and Late Differentiated CD8 T cell pool (CD45RA+CD27-). Of note, polyfunctional CD4 helper responses directed against tetanus toxoid and Survivin were induced by DPX-Survivac and possibly contributed to the maintenance of the CD8 T cell response. Taken together, the data presented herein shows that DPX-Survivac induced durable polyfunctional CD4 helper and CD8 T cells providing a strong rationale for combining the targeted immune therapy with immune modulation using metronomic cyclophosphamide.

Citation Format: Yoav Peretz, Dominike Sauvé, Dominic Gagnon, Salim Ahmed Khan, Geneviève Lévesque, Nathalie Saha, Gilbert Croteau, Valérie Hébert, Karyne Savard, Bader Yassine Diab, Jean-Francois Poulin, Claire Landry, Mohan Karkada, Marc Mansour. Immune profiling of patients vaccinated with the survivin targeted therapeutic vaccine DPX-Survivac demonstrates durable polyfunctional CD4+ and CD8+ T cells in ovarian cancer patients. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr CT411. doi:10.1158/1538-7445.AM2014-CT411
Abstract DDT02-01: In vitro and in vivo characterization of a novel anti-fibroblast growth factor receptor (FGFR) 2 antibody (BAY 1179470) for the treatment of gastric cancer
Authors: Kopitz Charlotte , Sommer Anette , Hammer Stefanie , Harrenga Axel , Stelte-Ludwig Beatrix , Dittmer Frank , Reetz Frank , May Ekkehard , Zierz Ruprecht , Wittemer-Rump Sabine , Schatz Christoph , Huynh H. T. , Ziegelbauer Karl , Kreft Bertolt
Published: May 2015

Gastric cancer is the second most common cause of cancer-related mortality worldwide, thus new treatment options are urgently needed. In a subset of gastric cancers, over-expression of fibroblast growth factor receptor 2 (FGFR2), a receptor tyrosine kinase, has been described and may represent a potential therapeutic target for the treatment of FGFR2-positive gastric cancer patients. To this end, we have generated a fully human anti-FGFR2 antibody (BAY 1179470) using the BioInvent Phage Display library. BAY 1179470 binds to a unique FGFR2-specific epitope that is present in all FGFR2 isoforms. Upon binding to FGFR2, BAY 1179470 induces receptor dimerization, internalization and degradation, resulting in significant tumor growth inhibition in vivo in cell line-based and patient-derived gastric cancer models overexpressing FGFR2. Additive anti-tumor efficacy in vivo was achieved by combining BAY 1179470 with either cisplatin or paclitaxel. BAY 1179470 is fully cross-reactive with FGFR2 orthologues of mouse, rat, pig, cynomolgus monkey and rhesus macaque. No significant safety findings have been seen in animal studies. Thus, BAY 1179470 represents a novel anti-FGFR2 antibody with high anti-tumor activity in gastric cancer models and an excellent preclinical safety profile. BAY 1179470 is currently being tested in a first-in-man study in all-comers (NCT01881217) in Japan.

Citation Format: Charlotte Kopitz, Anette Sommer, Stefanie Hammer, Axel Harrenga, Beatrix Stelte-Ludwig, Frank Dittmer, Frank Reetz, Ekkehard May, Ruprecht Zierz, Sabine Wittemer-Rump, Christoph Schatz, H. T. Huynh, Karl Ziegelbauer, Bertolt Kreft. In vitro and in vivo characterization of a novel anti-fibroblast growth factor receptor (FGFR) 2 antibody (BAY 1179470) for the treatment of gastric cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr DDT02-01. doi:10.1158/1538-7445.AM2014-DDT02-01
Abstract LB-139: IDH1 mutations alter citric acid cycle metabolism and increase dependence on oxidative mitochondrial metabolism
Authors: Grassian Alexandra R. , Parker Seth , Davidson Shawn , Divakaruni Ajit , Green Courtney , Zhang Xiamei , Slocum Kelly , Pu Minying , Lin Fallon , Vickers Chad , Joud-Caldwell Carol , Chung Franklin , Yin Hong , Handly Erika , Straub Christopher , Growney Joseph D. , Heiden Matt Vander , Murphy Anne , Pagliarini Raymond , Metallo Christian
Published: May 2015

Mutations in the genes encoding isocitrate dehydrogenase 1 and 2 (IDH1/2) occur in a variety of tumor types, resulting in production of the proposed oncometabolite, 2-hydroxyglutarate (2-HG). How mutant IDH alters central carbon metabolism, though, remains unclear. To address this question, we performed 13C metabolic flux analysis (MFA) on an isogenic cell panel containing heterozygous IDH1/2 mutations. We observe a dramatic and consistent decrease in the ability of IDH1, but not IDH2, mutant cell lines to utilize reductive glutamine metabolism via the carboxylation of α-ketoglutarate to isocitrate. Additionally we find that cells with IDH1 mutations exhibit increased oxidative tricarboxylic acid (TCA) metabolism. Similar metabolic trends were observed in vivo as well, and also in endogenous, non-engineered IDH1/2 mutant cell lines. Interestingly, IDH1-mutant specific inhibitors were unable to reverse the decrease in reductive metabolism, suggesting that this metabolic phenotype is independent of 2-HG. Furthermore, this metabolic reprogramming increases the sensitivity of IDH1 mutant cells to hypoxia or electron transport chain (ETC) inhibition in vitro. IDH1 mutant cells also grow poorly as subcutaneous xenografts within hypoxic in vivo microenvironments. These results suggest that exploiting metabolic defects specific to IDH1 mutant cells could be an interesting avenue to explore therapeutically.

Citation Format: Alexandra R. Grassian, Seth Parker, Shawn Davidson, Ajit Divakaruni, Courtney Green, Xiamei Zhang, Kelly Slocum, Minying Pu, Fallon Lin, Chad Vickers, Carol Joud-Caldwell, Franklin Chung, Hong Yin, Erika Handly, Christopher Straub, Joseph D. Growney, Matt Vander Heiden, Anne Murphy, Raymond Pagliarini, Christian Metallo. IDH1 mutations alter citric acid cycle metabolism and increase dependence on oxidative mitochondrial metabolism. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr LB-139. doi:10.1158/1538-7445.AM2014-LB-139
Abstract LB-154: Generation of antagonistic anti-DDR1 monoclonal antibodies as a potential novel cancer therapeutic
Authors: Jun H. Toni , Hynes Brandon , Macomber John L. , Lassen Andrew , Altobell Larry , MacLaren Josh , King David , Horlick Robert A.
Published: May 2015

Discoidin domain receptor 1 (DDR1) is a nonintegrin tyrosine kinase receptor for collagen implicated in diverse cellular roles, including cell adhesion, proliferation, and extracellular matrix remodeling. Multiple tumor types have been shown to overexpress DDR1, including lung, esophageal, breast, ovarian, and pediatric brain cancers, suggesting a potential role for DDR1 in tumor progression. Inhibition of DDR1 signaling may therefore represent a novel therapeutic approach for treating these cancers. Potential therapeutic molecules that modulate human DDR1 activity were identified by screening the AnaptysBio Evolvable Library (ABEL) of fully human germline antibodies. The initial ABEL screen used the soluble DDR-1 extra cellular domain, followed by secondary screening on cell-surface DDR-1. The resulting panel of human antibodies was affinity matured using SHM-XELTM which uses mammalian cell display of human IgG followed by in vitro somatic hypermutation (SHM) as described for a number of other antibodies (Bowers et al., 2014, Methods 65, 44-56). The matured antibodies bound specifically to DDR1 with high affinity (KD<1nM as measured by Biacore), and were able to functionally inhibit collagen triggered DDR1 signaling in multiple cell-based assays. The antibody panel shows selectivity for DDR1 over closely related molecules such as DDR2 and has other properties suitable for preclinical development, including cross-reactivity with the cynomolgus monkey DDR-1 ortholog and good biophysical properties. These antibodies are being further explored for their therapeutic efficacy in tumor types where DDR1 signaling plays a role in disease progression.

Citation Format: H. Toni Jun, Brandon Hynes, John L. Macomber, Andrew Lassen, Larry Altobell, Josh MacLaren, David King, Robert A. Horlick. Generation of antagonistic anti-DDR1 monoclonal antibodies as a potential novel cancer therapeutic. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr LB-154. doi:10.1158/1538-7445.AM2014-LB-154
Abstract LB-191: Astrocytes promote colonization of human brain with breast cancer cells via inhibition of KISS1 expression
Authors: Ulasov Ilya V. , Kaverina Natalya V. , Yoon JG , Lee Hwahyung , Sarvaiya Purvaba , Malin Dmitry , Cryns Vincent L. , Welch Danny R. , Cobbs Charles K.
Published: May 2015

Introduction: KISS1 metastatic suppressor is a negative regulator of tumor progression. Recently, we described that breast cancer metastatic cells to the brain exhibit much lower level of KISS1 expression detected by real time PCR and IHC. However little is known about the mechanism that regulates KISS1 in the brain metastatic cells and promotes brain metastases. We hypothesized that loss of KISS1 allows persistence of metastatic cells in the brain.

Results: Given the fact that some migrating breast cancer cells possess stem cells (CSC) properties, we stained the primary breast cancer and brain metastatic clinical specimens contain breast cancer cells using CD24, CD44, ESA1 and KISS1 markers. We observed that in the brain metastases, the CSC exhibit low/lack of KISS1 expression, whereas in the MDA-MB-231 model, blood derived circulating cells express comparative to primary xenografts level of KiSS1 expression. Then we co-cultured MDA-231-Br or CN34Br cells with either human microglia (HM) or human astrocytes (HA) to determine the conditions that affect KISS1 expression. We observed that primary human astrocytes downregulate the expression of KISS1, whereas HM conditional media had little effect. Furthermore, we detected that inhibition of KISS1 expression in breast metastatic cells is dependent on the expression of chemokines such CCL2 and CXCL12. This effect was dose dependent and required activation of cellular signaling in the tumor cells. Our data demonstrate that interaction of breast cancer cells with astrocytes inhibits KISS1 metastatic suppressor and contributes to tumorigenesis.

Conclusions: Our results provide a new insight into the mechanism that promotes brain metastases, and may have implications for the treatment and prevention of brain metastases.

Citation Format: Ilya V. Ulasov, Natalya V. Kaverina, JG Yoon, Hwahyung Lee, Purvaba Sarvaiya, Dmitry Malin, Vincent L. Cryns, Danny R. Welch, Charles K. Cobbs. Astrocytes promote colonization of human brain with breast cancer cells via inhibition of KISS1 expression. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr LB-191. doi:10.1158/1538-7445.AM2014-LB-191
Abstract LB-227: Preclinical pharmacology and clinical efficacy of AZD9150 (ISIS-STAT3Rx), a potent next-generation antisense oligonucleotide inhibitor of STAT3
Authors: Hong David , Kim Youngsoo , Younes Anas , Nemunaitis John , Fowler Nathan , Hsu Jeff , Zhou Tianyuan , Fayad Luis , Zhang Nancy , Piha-Paul Sarina , Woessner Richard , Nadella Murali V. , Lawson Deborah , Reimer Corinne , Jo Minji , Schmidt Joanna , Xiao Xiaokun , Greenlee Sarah , Hung Gene , Yamashita Mason , Blakey David C. , Monia Brett P. , Macleod A. Robert , Kurzrock Razelle
Published: May 2015

Genomic technologies have greatly expanded our understanding of cancer biology, although they have not yet been effectively translated into improved cancer therapeutics, partly due to the inability of available therapeutic modalities to target the most promising cancer driver pathways. In contrast to other therapeutic approaches, antisense technology allows the rational design of potent, sequence-specific inhibitors based on genome sequencing information alone. Recent human clinical data have demonstrated the potent activity of antisense oligonucleotides (ASOs) targeted to genes expressed by the liver. Here we describe preclinical and clinical activity of a high affinity, new generation chemistry (constrained ethyl modified) ASO, AZD9150. AZD9150 targets the mRNA coding for the transcription factor STAT3, which is considered a difficult protein to target therapeutically. Systemic delivery of unformulated AZD9150 resulted in strong inhibition of STA3 RNA and protein levels in demonstrates in a broad range of preclinical cancer models in vivo including several patient-derived xenograft models. AZD9150 administration resulted in 2 partial responses highly treatment-refractory diffuse large B-cell lymphoma and in 2 mixed responses in Hodgkin's Lymphoma patients in the phase I dose escalation study. These findings suggest that this technology has the potential to help bridge the pharmacogenomic divide in cancer drug discovery.

[D.H. and Y.K. contributed equally to this work.]

Citation Format: David Hong, Youngsoo Kim, Anas Younes, John Nemunaitis, Nathan Fowler, Jeff Hsu, Tianyuan Zhou, Luis Fayad, Nancy Zhang, Sarina Piha-Paul, Richard Woessner, Murali VP Nadella, Deborah Lawson, Corinne Reimer, Minji Jo, Joanna Schmidt, Xiaokun Xiao, Sarah Greenlee, Gene Hung, Mason Yamashita, David C. Blakey, Brett P. Monia, A. Robert Macleod, Razelle Kurzrock. Preclinical pharmacology and clinical efficacy of AZD9150 (ISIS-STAT3Rx), a potent next-generation antisense oligonucleotide inhibitor of STAT3. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr LB-227. doi:10.1158/1538-7445.AM2014-LB-227
Abstract LB-256: Immunotherapy using genetically modified T lymphocytes to target CLEC14A on the tumor vasculature
Authors: Zhuang Xiaodong , Kaul Baksho , Bentley Michael , Nagy Zsuzsanna , Giraudo Enrico , Bendle Gavin , Gilham David , Bicknell Roy , Lee Steven P.
Published: May 2015

Chimeric antigen receptor (CAR) therapy combines the specificity of a monoclonal antibody with the potent cytotoxic and immune regulatory activity of self-replicating T cells. This approach has shown remarkable efficacy in recent clinical trials targeting T cells to hematological cancers. More limited benefit has been reported with solid tumors, possibly because of lack of suitable target antigens, immune evasion mechanisms in the malignant cells and/or lack of T cell infiltration into the tumor tissue. An alternative approach that avoids some of these problems is to use a CAR that targets the tumor vasculature rather than the malignant cells directly.

CLEC14A is a glycoprotein recently identified to be selectively overexpressed on the surface of the tumor vasculature in a wide range of human cancers, including ovarian, liver, bladder, prostate, breast and kidney, and is therefore of considerable interest for tumor therapy. Here we explore for the first time the feasibility of targeting the tumor vasculature using CLEC14A-specific CARs. Initially we generated CARs from several CLEC14A-specific monoclonal antibodies and then expressed them in T cells. In vitro T cell functional assays demonstrated that these CARs can trigger potent antigen-specific activation including cytotoxicity in engineered T cells. These CARs recognize both human and mouse forms of CLEC14A, so safety/efficacy studies could be conducted using mouse models. Following infusion of escalating doses of engineered T cells into healthy mice, circulating CAR-T cells persisted for at least the next 5 weeks without signs of toxicity. To explore anti-tumor effects, Lewis lung carcinoma cells were subcutaneously implanted into C57BL6 mice and 4 days later mice received a single dose of CLEC14A-specific CAR-T cells. Our recent data demonstrate statistically significant inhibition of tumor growth in mice treated with CAR-T cells compared with mock-transduced T cells. The engineered cells also appeared to expand/accumulate at the tumor site. These data suggest CLEC14A can be safely and effectively targeted with CAR-T cells, potentially offering a potent and widely applicable cancer therapy.

Citation Format: Xiaodong Zhuang, Baksho Kaul, Michael Bentley, Zsuzsanna Nagy, Enrico Giraudo, Gavin Bendle, David Gilham, Roy Bicknell, Steven P. Lee. Immunotherapy using genetically modified T lymphocytes to target CLEC14A on the tumor vasculature. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr LB-256. doi:10.1158/1538-7445.AM2014-LB-256
Abstract LB-4: Ghrelin administration suppresses inflammation-associated colorectal carcinogenesis in mice
Authors: Kataoka Hiroaki , Kawaguchi Makiko , Kanemaru Ai , Fukushima Tsuyoshi , Matsumoto Nobuhiro , Nakazato Masamitsu
Published: May 2015

Background and aim: Ghrelin is a 28-amino-acid peptide, capable of stimulating growth hormone release. Because of its orexigenic effect, ghrelin is being developed as a therapeutic option for post-operative support and anorexia-cachexia syndrome of cancer patients. However, ghrelin has a multiplicity of physiological functions in various organs and immune system, and also affects cell proliferation. Therefore, it is important to know the effect of ghrelin administration on carcinogenesis and cancer progression in patients who are susceptible to cancer. This study aimed to test the effect of ghrelin on cancer promotion using murine intestinal carcinogenesis models.

Method: Effect of ghrelin knockout or exogenous ghrelin administration (3 nmoles/day, i.p.) on intestinal tumorigenesis was analyzed in two types of murine intestinal tumorigenesis models; ApcMin/+ mice (genetic cancer susceptibility model) and AOM/DSS-treated mice (inflammation-associated colon cancer model).

Results: The absence of ghrelin did not affect the number of intestinal tumors formed in ApcMin/+ mice and AOM/DSS-treated mice. Administration of ghrelin did not affect the number of intestinal tumors formed in ApcMin/+ mice, either. However, in AOM/DSS-treated mice, the administration of ghrelin significantly suppressed the formation of colon tumors (p < 0.001). The intestinal mRNA levels of inflammatory cytokines after DSS treatment were also suppressed by the administration of ghrelin.

Conclusion: Ghrelin administration suppresses inflammation-associated colorectal carcinogenesis, which is possibly mediated by anti-inflammatory effects of ghrelin.

Citation Format: Hiroaki Kataoka, Makiko Kawaguchi, Ai Kanemaru, Tsuyoshi Fukushima, Nobuhiro Matsumoto, Masamitsu Nakazato. Ghrelin administration suppresses inflammation-associated colorectal carcinogenesis in mice. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr LB-4. doi:10.1158/1538-7445.AM2014-LB-4
Abstract LB-92: Using immunohistochemistry to evaluate FGFR3 gene fusion recurrence and clinical associations in astrocytomas
Authors: Granberg Kirsi J. , Lehtinen Birgitta , Annala Matti J. , Haapasalo Hannu , Nykter Matti , Zhang Wei
Published: May 2015

Aberrant activation of fibroblast growth factor receptor 3 (FGFR3) has been reported to be tumorigenic in various cancer types. When FGFR3 is fused with another gene, most commonly with transforming acidic coiled coil 3 (TACC3), it can escape miRNA-mediated regulation and remain constitutively active most likely due to the dimerization via the domains of TACC3 or another fusion partner. Among brain cancers, gene fusions involving FGFR3 have been detected in glioblastoma and certain low grade gliomas. Furthermore, FGFR3-TACC3 fusion positive glioma cells have responded well to targeted therapies. The endogenous FGFR3 protein levels are extremely low in brain partly due to miR-99a-mediated regulation and gene fusions lead to prominent protein expression. In previous studies, FGFR3 fusion recurrence and fusion partners have been analyzed only in limited patient cohorts and there are no reports about their clinical associations. In this study, we have performed immunohistochemistry for FGFR3 on tissue microarrays with over 750 clinical astrocytoma samples (including grade 1-3 astrocytomas and glioblastomas). FGFR3 positive staining has been detected in subpopulations of all included tumor grades. The observed recurrences (1.8%-6.2%, depending on the grade) are consistent with previous sequencing-based studies. To validate immunohistochemistry-based approach for FGFR3 fusion detection, we are using frozen tumor material to confirm the presence of fusion with Western blotting and with PCR followed by Sanger sequencing. Furthermore, we have associated FGFR3 staining scores with clinical records for all the cases. FGFR3 staining is more common in females than males both in astrocytoma (7.6% vs 2.2%, pearson chi-square p-value 0.004, n=752) and glioblastoma cohorts (8.9% vs 2.1%, pearson chi-square p-value 0.008, n=494). In addition, FGFR3-staining negatively associates with aberrant p53 staining indicating p53 mutation (FGFR3-positive cases: 5.8% vs 1.8% in p53 low vs high groups, respectively, pearson chi-square p-value 0.030, n=399). Altogether, this study evaluates the suitability of immunohistochemical methods for FGFR3 fusion detection as well as investigates their recurrence and clinical significance in astrocytomas.

Citation Format: Kirsi J. Granberg, Birgitta Lehtinen, Matti J. Annala, Hannu Haapasalo, Matti Nykter, Wei Zhang. Using immunohistochemistry to evaluate FGFR3 gene fusion recurrence and clinical associations in astrocytomas. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr LB-92. doi:10.1158/1538-7445.AM2014-LB-92
Abstract SY07-03: Gain-of-function activities of mutant p53
Authors: Lozano Guillermina
Published: May 2015

Mutations in p53 in tumors vary from deletions and nonsense mutations that lead to the absence of p53 protein to missense mutations that disrupt the DNA binding function of p53 and that exhibit gain-of-function phenotypes. The p53R172H protein manifests one of its gain-of-function activities as a metastatic phenotype in vivo. To understand the contribution of mutant p53 to metastasis, expression arrays were used to compare primary osteosarcomas from p53R172H/+ mice with metastasis to osteosarcomas from p53+/- mice lacking metastasis. Of 213 gene differences, Pla2g16 was overexpressed in metastatic osteosarcomas and was of particular interest as it encodes a phospholipase that catalyzes phosphatidic acid into lysophosphatidic acid and free fatty acid (both implicated in metastasis). Functional analyses showed that Pla2g16 knockdown decreased migration and invasion in mutant p53 expressing cells, and vice versa, overexpression of Pla2g16 increased the invasion of p53-null cells. Mechanistically, various mutant p53 proteins bind the Pla2g16 promoter at ETS binding motifs and knockdown of ETS2 in particular suppressed mutant p53 induction of Pla2g16. Thus, our study identifies a phospholipase as a transcriptional target of mutant p53 that contributes to metastasis.

Citation Format: Guillermina Lozano. Gain-of-function activities of mutant p53. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr SY07-03. doi:10.1158/1538-7445.AM2014-SY07-03
Abstract 22: RECQ1 promotes the expression of genes significantly associated with cancer progression
Authors: Lu Xing , Li Xiao Ling , Parvathaneni Swetha , Lal Ashish , Sharma Sudha
Published: May 2015

RecQ helicases (represented by five homologs in humans: RECQ1, WRN, BLM, RECQ4, and RECQ5) are vital to maintain genomic stability under replication stress and germ line mutations in WRN, BLM and RECQ4 are associated with cancer predisposition syndromes. Expression of RECQ1 has been associated with the process of cellular transformation and over-expression of RECQ1 has been reported in a variety of human cancers. Furthermore, siRNA-mediated knockdown of RECQ1 has a cancer cell specific effect in cell culture models and also suppresses tumor growth in mouse xenografts including lung, liver, pancreatic and colorectal cancer. In a panel of 60 human cancer cell lines (NCI-60), we have found that RECQ1 mRNA expression positively correlates with the mesenchymal marker Vimentin and negatively correlates with epithelial markers such as KRT8. We also observed significant positive correlation of RECQ1 expression with Vimentin and epithelial to mesenchymal transition (EMT) promoting proteins and negative correlation with CDH1 (epithelial marker) in cancer patient samples from TCGA (The Cancer Genome Atlas). Importantly, quantitative real-time PCR analyses in a set of 16 colorectal cancer patient samples revealed strong positive correlation of RECQ1 expression with Vimentin. Furthermore, lower expression of RECQ1 significantly correlated with better prognosis of recurrence-free survival in breast and ovarian cancer patients. Global analyses of gene expression upon knockdown of RECQ1 in HeLa (cervical adenocarcinomas) and MDA-MB-231 (breast adenocarcinoma) cells identified a subset of genes involved in cell invasion, migration and adhesion as most robustly regulated by RECQ1. Consistent with this result, we demonstrate that knockdown of RECQ1 significantly inhibits cell migration and invasion. These findings identify a novel function of RECQ1 in regulation of gene expression and suggest that RECQ1 contributes to tumor development and progression, in part, through regulating the expression of key genes that promote invasion and metastasis of cancer cells. Additionally, our data suggests that RECQ1 expression may serve as predictive marker of clinical outcome.

Citation Format: Xing Lu, Xiao Ling Li, Swetha Parvathaneni, Ashish Lal, Sudha Sharma. RECQ1 promotes the expression of genes significantly associated with cancer progression. [abstract]. In: Proceedings of the AACR Special Conference: Cancer Susceptibility and Cancer Susceptibility Syndromes; Jan 29-Feb 1, 2014; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(23 Suppl):Abstract nr 22. doi:10.1158/1538-7445.CANSUSC14-22
Abstract IA18: Molecular basis for cancer susceptibility in Bloom's syndrome
Authors: Hickson Ian D.
Published: May 2015

BLM is a member of the RecQ family of DNA helicases, and is the protein defective in the cancer predisposition disorder, Bloom's syndrome. This autosomal recessive disorder is also associated with fertility, immune system and skin abnormalities. BLM forms part of an evolutionarily conserved complex of proteins including Topoisomerase IIIa, RMI1 and RMI2 (collectively called the BTRR complex). The BTRR complex has a well-defined role in S-phase, but our recent studies have uncovered a novel function in mitosis. All proliferating cells must faithfully segregate their newly replicated sister chromatids during mitosis. Any perturbation of this process, arising from either endogenous or exogenous sources, can lead to cell death or chromosomal instability. We have shown previously that common fragile site loci are difficult to segregate in anaphase and, as a result, create so-called ultra-fine anaphase bridges (UFBs). These bridges are present in virtually all cells in early anaphase and are decorated along their length by a SNF2 family protein, PICH, and the BTRR complex. We hypothesize that common fragile sites fail to complete replication in S-phase, which necessitates that action of the PICH and BTRR proteins to disentangle the incompletely replicated DNA in anaphase itself. We are analyzing these processes in yeast, chicken and human cells. For example, we have generated a PICH-deficient chicken DT-40 cell mutant, and have devised a system in yeast to generate a site-specific, artificial fragile site by blocking converging replication forks. Biochemically, we are analyzing the ability of purified PICH and BTRR proteins to disentangle a DNA substrate in vitro that mimics the structure that would form as two replication forks converge. We have also recently discovered a link between the action of DNA structure-specific nuclease, MUS81-EME1, and the frequency of fragile site expression and UFB formation. Results of these studies will be discussed.

Citation Format: Ian D. Hickson. Molecular basis for cancer susceptibility in Bloom's syndrome. [abstract]. In: Proceedings of the AACR Special Conference: Cancer Susceptibility and Cancer Susceptibility Syndromes; Jan 29-Feb 1, 2014; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(23 Suppl):Abstract nr IA18. doi:10.1158/1538-7445.CANSUSC14-IA18
Abstract A53: The specific FMS kinase inhibitor, DCC-3014, durably inhibits FMS kinase in vivo and blocks cancer bone invasiveness
Authors: Smith Bryan D. , Kaufman Michael D. , Leary Cynthia B. , Hood Molly M. , Lu Wei-Ping , Turner Benjamin A. , Vogeti Subha , Wise Scott C. , Flynn Daniel L.
Published: May 2015

Within the tumor microenvironment, tumor-associated macrophages (TAMs) rely on signaling through FMS kinase to promote tumor growth, angiogenesis, vasculogenesis, and metastasis. In addition, FMS-expressing macrophages communicate with the adaptive immune system, which can lead to cancer immunotolerance. Osteoclasts also depend on FMS kinase for differentiation and growth, and play a direct role in the ability of a tumor to metastasize to the bone and progress. Taken together, FMS kinase is an excellent target for small molecule therapy of tumor growth and bone metastasis. Using Deciphera Pharmaceuticals' approach to kinase inhibition, potent and specific FMS kinase inhibitors have been identified that achieve single digit nanomolar inhibition in biochemical and cellular assays of FMS activity. Due to their binding mode, these inhibitors retain potency in the presence of high ATP concentrations and also exhibit long off-rates from FMS kinase. Development candidate DCC-3014 is highly selective, inhibiting no other kinases within 100-fold of FMS potency, including related kinases KIT, PDGFRs, FLT3, and VEGFR2. DCC-3014 exhibits potent and durable inhibition of FMS in vivo in a pharmacokinetic/pharmacodynamic model. In a variety of cancer models, DCC-3014 or related analogs slow tumor growth, decrease tumor-promoting macrophages, and protect against osteolytic bone invasion. DCC-3014 is currently undergoing formal preclinical development.

Citation Format: Bryan D. Smith, Michael D. Kaufman, Cynthia B. Leary, Molly M. Hood, Wei-Ping Lu, Benjamin A. Turner, Subha Vogeti, Scott C. Wise, Daniel L. Flynn. The specific FMS kinase inhibitor, DCC-3014, durably inhibits FMS kinase in vivo and blocks cancer bone invasiveness. [abstract]. In: Abstracts: AACR Special Conference on Cellular Heterogeneity in the Tumor Microenvironment; 2014 Feb 26-Mar 1; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2015;75(1 Suppl):Abstract nr A53. doi:10.1158/1538-7445.CHTME14-A53
Abstract A69: MCT-1 expression promotes neoplastic multinucleation through the Src/p190B signaling activation in triple-negative breast cancer cells
Authors: Wu Meng-Hsun , Chen Yen-An , Chen Hsiao-Huei , Chang Ko-Wei , Chang I-Shou , Wang Lu-Hai , Hsu Hsin-Ling
Published: May 2015

Multinucleation is associated with malignant neoplasm; however, the molecular mechanism underlying the nuclear abnormality remains unclear. Loss or mutation of PTEN is frequently implicated in the development of malignancies. We now demonstrate that increased expression of the oncogene MCT-1 antagonizes PTEN gene expression, protein stability and activation, thereby further promoting PI3K/AKT signaling and survival rate of the PTEN-deficient cells. MCT-1 interacts with p190B and Src in the triple negative breast cancer cells, supporting they are in proximity of the signaling complexes in vivo. MCT-1 overexpression and PTEN loss synergistically augmented the Src/p190B signaling function leading to inhibition of RhoA activity. Under such condition, the incidence of mitotic catastrophes including spindle multipolarity and cytokinesis failure were enhanced, driving a Src/p190B-dependent neoplastic multinucleation. Reduced MCT-1 expression by shRNA dramatically repressed the Src/p190B function and improved the nuclear structure, as well as suppressed tumorigenicity of the PTEN-null triple negative breast cancer cells. Consistent with the oncogenic effects in vitro, clinical evidence has confirmed that MCT-1 and p190B genes are highly expressed in human breast cancers. Accordingly, MCT-1 gene induction is recognized as a potentially indicator of breast tumor development. Abrogating MCT-1 function may be a promising strategy for treatment of the triple negative breast cancer involving aberrant activation of Src and/or dysfunction of PTEN.

Citation Format: Meng-Hsun Wu, Yen-An Chen, Hsiao-Huei Chen, Ko-Wei Chang, I-Shou Chang, Lu-Hai Wang, Hsin-Ling Hsu. MCT-1 expression promotes neoplastic multinucleation through the Src/p190B signaling activation in triple-negative breast cancer cells. [abstract]. In: Abstracts: AACR Special Conference on Cellular Heterogeneity in the Tumor Microenvironment; 2014 Feb 26-Mar 1; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2015;75(1 Suppl):Abstract nr A69. doi:10.1158/1538-7445.CHTME14-A69
Abstract A72: Next-generation sequencing of disseminated tumor cells
Authors: Møller Elen K. , Kumar Parveen , Voet Thierry , Peterson April , Loo Peter Van , Mathiesen Randi , Fjelldal Renate , Grundstad Jason , Borgen Elin , Baumbusch Lars O. , Naume Bjørn , Børresen-Dale Anne-Lise , Nord Silje , Kristensen Vessela N.
Published: May 2015

Disseminated tumor cells (DTCs) detected in the bone marrow have been shown as an independent prognostic factor for women with breast cancer. However, the mechanisms behind the tumor cell dissemination are still unclear and more detailed knowledge is needed to fully understand why some cells remain dormant and others metastasize. Sequencing of single cells has opened for the possibility to dissect the genetic content of subclones of a primary tumor, as well as DTCs. Previous studies of genetic changes in DTCs have employed single-cell array comparative genomic hybridization which provides information about larger aberrations. To date, next generation sequencing provides the possibility to discover new, smaller and copy neutral genetic changes. In this study, we performed whole genome amplification and subsequently next generation sequencing to analyze DTCs from two breast cancer patients. We compared copy number profiles of the DTCs and the corresponding primary tumor generated from sequencing and SNP-CGH data, respectively. While one tumor revealed mostly whole arm gains and losses, the other had more complex alterations, as well as subclonal amplification and deletions. Whole arm gains or losses in the primary tumor were in general also observed in the corresponding DTC. Both primary tumors showed amplification of chromosome 1q and deletion of parts of chromosome 16q, which was recaptured in the corresponding DTCs. Interestingly, clear differences were also observed, indicating that the DTC underwent further evolution at the copy number level. This study provides a proof-of-principle for sequencing of DTCs and correlation with primary copy number profiles. The analyses allow insight into tumor cell dissemination and show ongoing copy number evolution in DTCs compared to the primary tumors.

Note: This abstract was not presented at the conference.

Citation Format: Elen K. Møller, Parveen Kumar, Thierry Voet, April Peterson, Peter Van Loo, Randi Mathiesen, Renate Fjelldal, Jason Grundstad, Elin Borgen, Lars O. Baumbusch, Bjørn Naume, Anne-Lise Børresen-Dale, Silje Nord, Vessela N. Kristensen. Next-generation sequencing of disseminated tumor cells. [abstract]. In: Abstracts: AACR Special Conference on Cellular Heterogeneity in the Tumor Microenvironment; 2014 Feb 26-Mar 1; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2015;75(1 Suppl):Abstract nr A72. doi:10.1158/1538-7445.CHTME14-A72