bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5138: The PD-1/PD-L1 axis and clinical outcomes in head and neck squamous cell carcinomas
Authors: Trivedi Sumita , Seethala Raja R. , Ferris Robert L.
Published: July 2016

Although novel therapeutic agents targeting PD-L1 and PD-1 have emerged, our understanding of the complex interaction between the tumor microenvironment and this checkpoint receptor-ligand axis remains incomplete. PD-L1 protein expression has been demonstrated by immunohistochemistry in several solid tumors including head and neck squamous cell carcinomas (HNSCC). However, the prognostic significance of PD-L1 overexpression in HNSCC remains inconclusive. Furthermore, the interaction between human papillomavirus (HPV) positive tumors, PD-L1 on tumor cells and PD-1 on immune cells has yet to be demonstrated in an architecturally intact tumor environment.

Here we demonstrate PD-L1 expression in a large cohort of mixed HPV+ and HPV-, surgically treated HNSCC using immunohistochemistry. Our findings confirm that more HPV+ tumors express PD-L1 compared with HPV- tumors (72% compared with 44%). Clinically, this correlates with improved overall and disease-free survival in in patients with HPV+ PD-L1+ tumors compared with HPV- PD-L1- tumors. By examining the intact tumor, we visualized distinct patterns of PD-L1 staining, either peripherally, at the tumor front or non-peripherally. Tumors with a peripheral staining pattern showed a trend towards decreased overall and disease-free survival, compared to those with a non-peripheral pattern of staining. By double staining PD-L1 and PD-1, we analyzed the co-expression of the ligand on tumor cells and its receptor on neighboring immune cells. Interestingly, patients whose tumors displayed co-localization of PD-1 and PD-L1 showed a trend towards improved disease-free survival compared to tumors where there was no co-localization of PD-1 and PD-L1.

In conclusion, out data suggest that the presence of PD-L1 and PD-1 alone may not be sufficient to guide clinical treatment rather; careful examination of the intact tumor microenvironment may inform more targeted therapy.

Citation Format: Sumita Trivedi, Raja R. Seethala, Robert L. Ferris. The PD-1/PD-L1 axis and clinical outcomes in head and neck squamous cell carcinomas. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5138.
Abstract 5148: Angiotensin II type I receptor drives cancer progression through the FLJ10540/S100A9 pathway in oral cancer
Authors: Chen. Chang-Han
Published: July 2016

Oral cancer is the eighth most common cancers and a growing problem worldwide. Despite the easy approach for visual examination, oral cancer is often detected at advanced stages leading to severely decreased patient survival. Identifying reliable molecular markers for oral cancer provided in developing new therapeutic interventions is urgently needed. In this study, we explored that angiotensin II type I receptor (AT1R) was overexpression in tumor tissues of oral cancer patients, and its expression also significantly correlated with advanced TNM-stage, lymph node stage, and the poor 5-year survival rate. Functionally, gain-of function of AT1R in oral cancer cells promoted tumor cell growth in vitro and in xenograft model by MTT/colony formation/BrdU incorporation/time-lapse/IVIS, and 3D-Sonographic imaging appraoches. Furthermore, using the same cell panels, the migratory and invasive abilities were increased by wound healing and Transwell assays and the epithelial-mesenchymal transition (EMT) markers, such as N-cadherin, fibronectin and vimentin were also elevated. Conversely, loss-of function of AT1R in oral cancer cells by using siRNA and CRISPR/Cas9 systems demonstrated that the abilities of tumor proliferation, and motility were decreased in vitro and in vivo. By using syn-expression concept combinging with bioinformatic appraoch, we found that FLJ10540 was a novel dwonstream target of AT1R. FLJ10540 expression was not only regulated by AT1R, but also participated in AT1R-elicited cancer progression in vitro and in vivo. We established FLJ10540 knockout oral cancer cell lines by CRISPR/Casp9 to perform DNA microarray and found that S100A9 was a top ranking target among the differrentially expression in down-regulated candidate sets. By using biochemical approaches and xenograft model, the post-transcriptional level of S100A9 was modulated by FLJ10540. In addition, the AT1R-elicited cancer progression was required for FLJ10540/S100A9 expression. Finally, immunohistochemical analysis of human aggressive oral cancer specimens showed a significant positively correlation between AT1R/FLJ10540 and S100A9 expression. Taken together, these findings suggest that AT1R is not only an important prognostic factor but also a new therapeutic target in the FLJ10540/S100A9 pathway for human cancer.

Citation Format: Chang-Han Chen. Angiotensin II type I receptor drives cancer progression through the FLJ10540/S100A9 pathway in oral cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5148.
Abstract 5165: ADAM17: a common effector for RAS and MET-driven transformation of primary keratinocytes
Authors: Cataisson Christophe , Klosterman Mary , Shibuya Kelly , Merlino Glenn , Yuspa Stuart H.
Published: July 2016

The purpose of the current study is to determine the contribution of the metalloproteinase ADAM17 (a disintegrin and metalloprotease 17) to the transformation driven by activated MET or oncogenic RAS in primary keratinocytes. Transgenic keratinocytes overexpressing HGF (MT-HGF) to activate MET and keratinocytes transduced with an oncogenic RAS share identical phenotypic and biochemical features of transformation and produce squamous tumors in vivo. In both keratinocyte populations, these common features arise from autocrine activation of EGFR through elevated expression and release of EGFR ligands. Amphiregulin (AREG) blockade in MT-HGF keratinocytes decreases MET-mediated EGFR transactivation. Inhibition of EGFR ablates the initiated signature of MT-HGF keratinocytes in vitro and causes regression of tumors from MT-HGF keratinocytes tranplanted in vivo. Deletion or knock-down of ADAM17 decreases EGFR activation in both RAS and MT-HGF keratinocytes and reverses the transformation associated gene signature in RAS keratinocytes. Using AREG release as an indicator of ADAM17 activity, we determined that knock-down of either the SRC kinase or inactive Rhomboid 2 (iRhom2) reduces the ability of keratinocytes to release AREG upon HGF stimulation. Collectively our data suggest that MET-mediated transformation of keratinocytes occurs through EGFR transactivation and that ADAM17 is a necessary effector for RAS and MET-driven transformation.

Citation Format: Christophe Cataisson, Mary Klosterman, Kelly Shibuya, Glenn Merlino, Stuart H. Yuspa. ADAM17: a common effector for RAS and MET-driven transformation of primary keratinocytes. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5165.
Abstract 5166: Gene expression microarray analysis identifies association of genes in the integrin/fak signaling pathway with claudin-7 in human colon cancer tissue
Authors: Kong Lu , Ding Lei
Published: July 2016

The role and mechanism of the integrin/focal adhesion kinase (FAK)/ERK signaling pathway in the progression of many malignant tumor types, including colon cancer, have been widely reported. The claudin family of proteins is integral to the structure and function of tight junctions. The role of claudin-7 (Cldn-7, CLDN7) in the regulation of the integrin/FAK/ERK signaling pathway remains poorly understood. Therefore, we investigated differences in gene expression, primarily focusing on CLDN7 and integrin/FAK/ERK signaling pathway genes, in colon cancer and adjacent normal tissues. Quantitative real-time RT-PCR verified the results for mRNA expression, and immunohistochemistry verified the results for protein expression. In silico analysis was used to predict co-regulation between Cldn-7 and integrin/FAK/ERK signaling pathway members. Our results showed marked cancer stage-specific decreases in the protein expression of Cldn-7, Gelsolin, MAPK1 and MAPK3 in colon cancer samples. The observed changes in the expression of all proteins except Cldn-7 were in agreement with the changes in the corresponding mRNA levels. Cldn-7 was predicted not to directly regulate any genes in the integrin/FAK/ERK signaling pathway, but these factors may participate in a common network that regulates cancer progression, in which the MAPK pathway is the central node.

Citation Format: Lu Kong, Lei Ding. Gene expression microarray analysis identifies association of genes in the integrin/fak signaling pathway with claudin-7 in human colon cancer tissue. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5166.
Abstract 5170: Snail-dependent ESRP1 silencing drives malignant conversion of human pulmonary epithelial cells
Authors: Walser Tonya C. , Jing Zhe , Tran Linh , Lin Ying , Zhu Li , Sharma Sherven , Ooi Aik , Gomperts Brigitte N. , Shay Jerry W. , Larsen Jill E. , Gao Boning , Minna John D. , Fishbein Micihael C. , Dubinett Steven M.
Published: July 2016

The significance of Snail expression in the early pathogenesis of non-small cell lung cancer has not been determined. Here, we report robust Snail expression in premalignant lung lesions relative to histologically normal-appearing pulmonary and lung cancer tissues. Utilizing immortalized human bronchial epithelial cells (HBECs) to isogenically model the contribution of the transcriptional repressor Snail to lung carcinogenesis, we document Snail-dependent anchorage-independent growth of pulmonary epithelial cells in vitro, as well as their transformation, primary tumor growth, and metastatic behavior in vivo. We delineate ESRP1 tumor suppressor silencing as a requirement for the Snail-driven transformation and identify ESRP1 silencing in Snail-expressing pulmonary premalignant lesions in situ. Utilizing this HBEC lung carcinogenesis model, we determined that Snail drives the expansion and malignant conversion of an ALDH+CD44+CD24- subset of pulmonary stem cells. Apoptosis-resistance and expression changes in oncogenes and tumor suppressors were noted, and ESRP1 was identified as a mediator of the malignant conversion of these Snail-primed pulmonary. Collectively, this is the initial report of a Snail-ESRP1-cancer axis that is operative during lung carcinogenesis and the first characterization of potentially targetable mediators of this normal to cancer continuum.

Citation Format: Tonya C. Walser, Zhe Jing, Linh Tran, Ying Lin, Li Zhu, Sherven Sharma, Aik Ooi, Brigitte N. Gomperts, Jerry W. Shay, Jill E. Larsen, Boning Gao, John D. Minna, Micihael C. Fishbein, Steven M. Dubinett. Snail-dependent ESRP1 silencing drives malignant conversion of human pulmonary epithelial cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5170.
Abstract 5173: Role of Gli2 in mediating the progression of prostate cancer to CRPC
Authors: Xia Lu , Qin Tai , Sun Luzhe
Published: July 2016

Background: Prostate cancer is a leading cause of cancer-related death in the US men, largely due to the development of castration resistant prostate cancer (CRPC) in the patients treated with androgen deprivation therapy. There is increasing evidence indicating that Hedgehog pathway is involved in the malignancy of prostate cancer, which includes the finding of two prostate tumors with loss-of-function mutations in the SuFu gene, correlation of tumor grade with high expression of Sonic hedgehog, Gli2 and/or hedgehog target genes including Ptch1 and HIP, and increased Gli2 expression in therapy-resistant tumor cells from patients. In addition, treatment of mice with hedgehog inhibitors significantly attenuated the growth of prostate tumor xenografts. Some recent studies also implicated a crosstalk between Hedgehog/Gli signaling and androgen signaling pathways in prostate cancer. However, the role of Gli2 in CRPC progression has not been investigated. Methods: A human Gli2 specific small hairpin RNA (shRNA) expression vector was constructed using Tet-PLKO-puro lentivector, which was delivered into the androgen-dependent human prostate cancer LNCaP cells. RT-PCR, Western Blot and Luciferase Reporter Assay were performed to confirm doxycycline-inducible Gli2 knockdown. The effect of Gli2 knockdown on anchorage-dependent or -independent growth was assessed with MTT and Soft Agar Assays respectively. Cells were cultured in androgen depleted medium or treated with the anti-androgen, Casodex, to mimic androgen deprivation condition. Mouse xenograft models were used to assess the role of Gli2 in the development of CRPC in vivo. Results: Androgen deprivation promoted the expression of Hedgehog signaling components including Gli2 in the LNCaP cells. Gli2 knockdown inhibited anchorage-dependent and -independent growth of LNCaP cells. Casodex complemented the inhibition of Gli2 knockdown on the growth of LNCaP cells. Gli2 knockdown prevented the outgrowth of androgen-independent cells from LNCaP cells cultured in androgen depleted medium. Induced-knockdown of Gli2 by doxycycline inhibited the growth of castration-resistant tumors in vivo in mice. Conclusions: Our study demonstrated the important role of Gli2 in the development of CRPC. Therefore, it may have the potential to serve as a novel therapeutic target for the prevention of CRPC.

Citation Format: Lu Xia, Tai Qin, Luzhe Sun. Role of Gli2 in mediating the progression of prostate cancer to CRPC. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5173.
Abstract 5184: MiXeno Humanized Mouse Model as an Efficient Tool to Evaluate in Vivo Activity of Anti-Cancer Immunotherapeutics
Authors: Zhang Juan , Qiao Meng , Ding Jian , Wang Zhensheng , Li zhongliang , Shi Qian
Published: July 2016

Background: Fighting cancer by taking advantage of a patient's own immune system is showing considerable success in oncology research. Targeting therapeutic antibodies against tumor-associated antigens or the host immune system has been shown to prolong survival of patients in various tumor types. However, the lack of experimental immunotherapy models is a major obstacle toward better evaluating new immunotherapeutics and testing their exploratory combination strategies. To address this need, we set out to validate a new platform MiXenoTM by implanting human PBMC to immune-deficient mice. The MiXenoTM mice were partially reconstituted with human immune components. In vivo efficacy of several immunotherapeutics were evaluated.

Results: We have reported before that several MiXenoTM models could be used to evaluate anti-human PD-1 or PD-L1 antibodies, where human T cells were well reconstituted in the mouse system. Here one of our MiXenoTM collection: HCC827 model was further optimized and identified as a PD-1/PD-L1 inhibitor highly sensitive model. Involvement of T cells in the tumor microenvironment was observed in HCC827 tumors. Furthermore, the dynamics of NK cell population in blood, tumor and other organs (e.g. spleen and lymph nodes) was established in MiXenoTM models. NK cell function was evaluated as well.

Conclusion: The MiXenoTM model provides an alternative to the full stem cell reconstitution approach, and may allow various types of immunotherapies being evaluated in these partially humanized xenograft models.

Citation Format: Juan Zhang, Meng Qiao, Jian Ding, Zhensheng Wang, zhongliang Li, Qian Shi. MiXeno Humanized Mouse Model as an Efficient Tool to Evaluate in Vivo Activity of Anti-Cancer Immunotherapeutics. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5184.
Abstract 5189: Caloric restriction augments the chemotherapeutic response in a murine triple negative breast cancer model
Authors: Simone Brittany , Dan Tu , Palagani Ajay , LaRose Meredith , Savage Jason E. , Simone Nicole
Published: July 2016

Purpose:

Triple negative breast cancers (TNBC) are a highly aggressive subtype of breast cancer, linked with a poor prognosis. TNBC tumors have a higher propensity for metastasis and unfortunately have fewer treatment options due to their lack of estrogen, progesterone or Her-2/neu receptors. We sought to determine if caloric restriction (CR) could augment the response of chemotherapy including docetaxel and cisplatin in a TNBC model and improve survival.

Methods:

An orthotopic mouse model using a highly metastatic, luciferase-tagged TNBC cell line (4T1), was used to generate palpable tumors. Mice were then treated with CR, cisplatin alone, docetaxel alone or a combination of CR and chemotherapy. Three times a week, primary tumors were measured with calipers and in vivo imaging was performed to evaluate the metastatic burden of disease. Molecular evaluation of the tumors was performed, generating a mechanistic hypothesis for increased sensitivity to chemotherapy.

Results:

Combining either docetaxel or cisplatin with CR, significantly decreased the rate of primary tumor growth (p<0.001). Average time to metastases (19 days in ad lib group) was increased by an average of 10.5 +/- 3 days with the addition of either docetaxel or cisplatin, and increased by an additional 6 days in the CR+cisplatin (35 days until metastases, p<0.01). The addition of CR to chemotherapy decreased the overall number and volume of lung metastases, and increased overall survival as compared with those groups receiving chemotherapy alone. Molecularly, CR was noted to augment the effect of chemotherapy by decreasing inflammation that results from IL-1β mediated acute RT toxicity and molecular effectors in the insulin (insulin, IGF-1, IGF-BP3 and IGF-1R) and adipokine signaling pathways (decreased leptin and increased adiponectin).

Conclusions:

CR augments the effect of systemic chemotherapy in an aggressive TNBC mouse model, in part by decreasing the inflammatory response. The novel use of CR in combination with systemic chemotherapy has the potential to enhance clinical outcomes for patients with triple negative breast cancer and should be tested in the clinical setting.

Citation Format: Brittany Simone, Tu Dan, Ajay Palagani, Meredith LaRose, Jason E. Savage, Nicole Simone. Caloric restriction augments the chemotherapeutic response in a murine triple negative breast cancer model. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5189.
Abstract 5190: Correspondence between clinical patient outcome and in vivo carboplatine response of high grade ovarian cancer PDX
Authors: du Manoir Stan P. , Delpech Helene , Orsetti Beatrice , Tessier Jeremy , Colombo Pierre-Emmanuel , Theillet Charles G.
Published: July 2016

Survival rate for high grade ovarian cancer (HGOC) remains low and has not significantly evolved for several years. This poor outcome is due to several mechanisms of resistance (initial or acquired) in different context (Homologous recombination deficience, CCNE1 amplification). We developed a collection of High-grade ovarian cancer Patient Derived Xenografts (PDX) with extensive characterization (CGH, transcriptome and exome sequencing, Colombo et al, Oncotarget. 2015 Sep 29;6(29):28327-40). Our objective is to create a panel of HGOC PDX which encompasses main types of platinium sensitivity/resistance in HGOC. We selected 10 PDX models (9 serous histiotypes and one carcinosarcomas) among our collection of 39 HGOC PDXs from two patient extreme platinium response categories (resistant with a progression-free inferior to 6 months and sensible with a progression-free higher than 18 months, some of the sensitives with BRCA1 mutations). These PDXs included PDXs were grafted sub-cutaneously on nude mice and submitted to carboplatin treatment (50mg/kg twice a week, for 4 weeks) and impact on tumor growth was evaluated as well as histological characterization. For the carcinosarcomas, no selection of the epithelial or sarcomatous component upon treatment was observed. Outgrowth of the tumors after end of treatment is currently under assessment.

Remarkably, tumor PDXs response was in accordance with clinical status with complete and partial response for the sensitive PDXs; slowed growth under treatment or stabilization for resistant PDXs. Outgrowth of the tumor after the end of treatment is currently under assessment.

For each PDXs model, the large majority of the 7-9 treated tumor reacted in identical fashion if classified according to RECIST criteria with occasionally divergent responses reflecting tumor heterogeneity. Genetic and histological characterization of these outliers is ongoing.

This collection is a valuable resource to model carboplatine resistance in a well-defined context and will be the cornerstone of our future studies to conduct preclinical drugs tests, in particular for combination selected in vitro and to evaluate clonal evolution under treatment.

Citation Format: Stan P. du Manoir, Helene Delpech, Beatrice Orsetti, Jeremy Tessier, Pierre-Emmanuel Colombo, Charles G. Theillet. Correspondence between clinical patient outcome and in vivo carboplatine response of high grade ovarian cancer PDX. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5190.
Abstract 5216: Broadening tumor spectrum in Lynch syndrome: increased incidence for 15 distinct cancer types
Authors: Therkildsen Christina , Ladelund Steen , Lindberg Lars Joachim , Smith-Hansen Lars , Nilbert Mef
Published: July 2016

Introduction: Lynch syndrome is a multi-tumor syndrome predominantly associated with an increased risk of colorectal cancer, but is indeed linked to a yet uncertain number of extra-colonic tumor types.

Methods: In order to determine the risk for extra-colonic tumors in Lynch syndrome, we determined the incidence rates for 21 distinct malignancies in the national Danish Lynch syndrome cohort compared to an age-and sex matched control cohort from the Danish population.

Results: Based on data from 1494 Lynch syndrome mutation carriers, significantly increased risks were demonstrated for 15 cancer types. These included Lynch syndrome-related malignancies, such as cancer of the endometrium, the ovary, the small bowel, the urinary tract and brain tumors as well as tumor types with an undefined role in Lynch syndrome, such as breast cancer, gastric cancer, pancreatic cancer, skin cancer, prostate cancer, lung cancer, kidney cancer, and testicular cancer. Distinctive peak ages were defined with the highest incidence rate ratio at age 30-49 years for ovarian cancer and 50-69 years for endometrial cancer, breast cancer, and brain tumors to continuously increasing incidence rate ratios with increasing age for e.g. urothelial cancer, gastric cancer, pancreatic cancer and skin cancer.

Conclusion: The broad tumor spectrum in Lynch syndrome is of relevance to consider during genetic counseling, whereas the variable peak incidence ages provide a basis for precision surveillance programs that consider the different risks throughout life.

Citation Format: Christina Therkildsen, Steen Ladelund, Lars Joachim Lindberg, Lars Smith-Hansen, Mef Nilbert. Broadening tumor spectrum in Lynch syndrome: increased incidence for 15 distinct cancer types. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5216.
Abstract 5220: Evaluation of ACMG guideline classified variants in 180 cancer and incidental non-cancer genes in families with breast/ovarian cancer
Authors: Maxwell Kara N. , Hart Steven N. , Vijai Joseph , Schrader Kasmintan A. , Thomas Tinu , Wubbenhorst Bradley , Ravichandran Vignesh , Moore Raymond M. , Hu Chunling , Guidugli Lucia , Wenz Brandon , Slavin Thomas P. , Domchek Susan M. , Robson Mark E. , Szabo Csilla , Neuhausen Susan L. , Weitzel Jeffrey N. , Offit Kenneth , Couch Fergus J. , Nathanson Katherine L.
Published: July 2016

Sequencing tests assaying panels of genes or whole exomes are widely available for cancer risk evaluation. However, methods for classification of variants resulting from this testing are not well studied. We evaluated the ability of American College of Medical Genetics and Genomics (ACMG) guidelines to define the rate of mutations and variants of uncertain significance (VUS) in 180 medically relevant genes, including all ACMG designated reportable cancer and non-cancer genes, in individuals who met guidelines for hereditary cancer risk evaluation. We performed whole exome sequencing in 404 individuals in 257 families and classified 1640 variants from these genes. Potentially clinically actionable (likely-pathogenic/pathogenic, LP/P) versus nonactionable (VUS/likely-benign/benign) calls were 92% and 88% concordant with locus specific databases and Clinvar, respectively. LP/P mutations were identified in 11 of 25 breast cancer susceptibility genes in 27 BRCA1/2 negative families (11%). Evaluation of 84 additional autosomal dominant cancer susceptibility genes identified LP/P mutations in only four additional families (1.7%), suggesting they do not influence risk in this cohort. However, individuals from nine of 257 families (3.5%) had incidental LP/P mutations in 32 non-cancer disease genes, and 7% of individuals were monoallelic carriers of an LP/P mutation in 39 autosomal recessive cancer syndrome genes. Furthermore, 90% of individuals had at least one VUS. In summary, these data support the clinical utility of ACMG variant classification guidelines. In addition, evaluation of extended panels of cancer genes in breast/ovarian cancer families leads to only an incremental clinical benefit but substantially increases the complexity of the results.

Citation Format: Kara N. Maxwell, Steven N. Hart, Joseph Vijai, Kasmintan A. Schrader, Tinu Thomas, Bradley Wubbenhorst, Vignesh Ravichandran, Raymond M. Moore, Chunling Hu, Lucia Guidugli, Brandon Wenz, Thomas P. Slavin, Susan M. Domchek, Mark E. Robson, Csilla Szabo, Susan L. Neuhausen, Jeffrey N. Weitzel, Kenneth Offit, Fergus J. Couch, Katherine L. Nathanson. Evaluation of ACMG guideline classified variants in 180 cancer and incidental non-cancer genes in families with breast/ovarian cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5220.
Abstract 523: Activating mutations in EGFR abrogate hypoxia-associated radiation resistance in non-small cell lung cancer
Authors: Saki Mohammad , Makino Haruhiko , Tomimatsu Nozomi , Ding Lianghao , Story Michael D. , Burma Sandeep , Nirodi Chaitanya
Published: July 2016

Hypoxic tumors tend to be 3-5 times more resistant to ionizing radiation relative to well-oxygenated tumors in part because oxygen is required to stabilize radiation-induced DNA damage. Tumor-adaptive responses also contribute to hypoxia-associated radioresistance but the precise mechanisms are not fully understood. Here we test our evidence-based hypothesis that the epidermal growth factor receptor (EGFR), which is frequently over-expressed in non-small cell lung carcinomas (NSCLCs), has critical roles in a tumor adaptive DNA damage response (DDR) pathway in hypoxic tumors. We find that, despite elevated EGFR activity and pro-survival signaling, hypoxia-associated radioresistance is dramatically reduced in a biologically distinct class of NSCLCs harboring activating mutations, L858R and ΔE746-E750 in EGFR. Hypoxic NSCLCs with EGFR mutations exhibit a unique spectrum of basal and hypoxia-induced DSBs not found in hypoxic wild type EGFR-expressing NSCLCs. These lesions dominate the S-phase of the cell cycle and are linked to collapsed replication forks in hypoxic cells. Replication fork-associated DSBs are repaired in wild type, but not in mutant, EGFR expressing NSCLCs. Microarray gene expression analysis revealed that mutant, but not wild type, EGFR expression results in an altered DDR characterized by dramatic down-regulation of several key components of the HR pathway within 24 hours of exposure to 0.1% oxygen. Our model predicts that NHEJ, not HR, dominates an altered DDR in hypoxic tumors. Elevated EGFR signaling may contribute to this altered DDR state where the EGFR-NHEJ axis becomes critical for hypoxia associated radioresistance and could be a potential target for hypoxic tumor radiosensitization.

Citation Format: Mohammad Saki, Haruhiko Makino, Nozomi Tomimatsu, Lianghao Ding, Michael D. Story, Sandeep Burma, Chaitanya Nirodi. Activating mutations in EGFR abrogate hypoxia-associated radiation resistance in non-small cell lung cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 523.
Abstract 5240: Lacking hepatic retinoid storage in mice suppresses NASH-related hepatocarcinogenesis
Authors: Ideta Takayasu , Shirakami Yohei , Miyazaki Tsuneyuki , Sakai Hiroyasu , Tanaka Takuji , Blaner William S. , Shimizu Masahito
Published: July 2016

Non-alcoholic fatty liver disease (NAFLD), which is strongly associated with metabolic syndrome, has become one of the most common causes of chronic liver disease in developed countries. Some patients with NAFLD develop a more serious form of the disease, non-alcoholic steatohepatitis (NASH), and NASH can develop cirrhosis or even hepatocellular carcinoma (HCC). Although hepatic retinoid (vitamin A) stores are progressively lost during the development of liver diseases, how this affects NAFLD/NASH and NASH-related hepatocarcinogenesis is unknown. In order to investigate this, we used streptozotocin (STZ) and high-fat diet (HFD) to induce NASH and related hepatic tumorigenesis in matched wild-type and lecithin:retinol acyltransferase (LRAT) knockout (KO) mice, which lack stored retinoid in the liver. LRAT is the sole enzyme responsible for hepatic retinyl ester synthesis since LRAT KO mice completely lack lipid droplets in hepatic stellate cells. Mice were injected with STZ within 48 hours after birth and then fed HFD from 4 to 16 weeks of age. At the termination of the experiment, liver tumors were observed macroscopically and microscopically in both groups. In LRAT KO mice, the development of hepatic adenoma and HCC was significantly suppressed compared to control group. LRAT KO mice showed decreased expression levels of cyclin D1 and TGF-beta mRNA in the liver. The serum levels of d-ROMs and BAP were measured and the d-ROM/BAP ratio, which indicates oxidative stress, markedly decreased in LRAT KO mice. The fibrosis in the liver of the LRAT KO mice was decreased. In addition, liver fibrosis in LRAT KO mice was suppressed. These findings indicate that LRAT KO mice are less susceptible to STZ and HFD-induced liver tumorigenesis due to regulation of cell cycle and attenuation of oxidative stress.

The amount of hepatic retinoid may affect the progression of NASH and the development of NASH-related HCC.

Citation Format: Takayasu Ideta, Yohei Shirakami, Tsuneyuki Miyazaki, Hiroyasu Sakai, Takuji Tanaka, William S. Blaner, Masahito Shimizu. Lacking hepatic retinoid storage in mice suppresses NASH-related hepatocarcinogenesis. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5240.
Abstract 5264: The National Cancer Institute's biospecimen research database: A community resource for improving specimen quality
Authors: Engel Kelly B. , Greytak Sarah R. , Guan Ping , Moore Helen
Published: July 2016

Tissue and blood biospecimens used in cancer research are collected, processed and stored using standard operating procedures (SOPs) that can vary tremendously across institutions. This lack of standardization in biospecimen collection and processing practices across sample sets and institutions can be a contributing source of research variability and can compromise analytical results. In an effort to minimize preanalytical variability and improve the quality of biospecimens used for cancer research, the National Cancer Institute's Biorepositories and Biospecimen Research Branch (BBRB) developed the Biospecimen Research Database (BRD; http://biospecimens.cancer.gov/brd). The BRD is a free and publically accessible online database that aims to (i) improve access to peer-reviewed articles that investigate the impacts that biospecimen collection and handling practices can have on clinical, molecular, and proteomic endpoints, (ii) facilitate transparency between institutions and researchers by promoting SOP sharing and distribution, and (iii) improve the quality of biospecimens through the development of evidence-based procedural guidelines. To date, the BRD houses more than 2,300 articles and 200 SOPs from 30 different participating biobanking institutions. Articles are meticulously categorized and annotated by a team of Ph.D.-level scientists according to the type of biospecimen and technology platform used and the preanalytical factors investigated. Literature contained within the BRD serves as the foundation for internally developed reviewed papers as well as evidence-based procedural guidelines, termed Biospecimen Evidence-Based Practices, which focus on biospecimen collection, preservation, and processing. In addition, more than 90% of the BRD's SOP library was contributed by other government offices, non-profit, private, and international biobanks and institutions. BBRB has invested a substantial and continued level of effort in the establishment of this scientifically accurate and robust database. Ongoing participation by the scientific community in the form of public commenting, article suggestions and SOP submissions is encouraged and will increase both the diversity and value of the BRD.

Citation Format: Kelly B. Engel, Sarah R. Greytak, Ping Guan, Helen Moore. The National Cancer Institute's biospecimen research database: A community resource for improving specimen quality. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5264.
Abstract 5275: An integrated pipeline for TCGA data analysis
Authors: Yu Kaixian , Xu Yun , Tang Ke , Steppi Albert , Zhou Jun , Ouyang Zheng , Zhang Jinfeng
Published: July 2016

The accumulation of publicly available high-throughput experimental data has made analyzing the data a bottleneck in scientific discovery. In this study, we explore a computational, high-throughtput approach for identifying cancer related biological factors using the data available at the Cancer Genome Atlas (TCGA). We developed an integrative genomic analysis (IGA) tool, which performs multiple analysis tasks, including gene/protein level differential expression analysis, gene set level enrichment analysis and network/pathway level comparison. Several data types can be analyzed, including RNA-seq, DNA methylation, miRNA-seq, and proteomics. By varying cancer types, race groups, and data types, we are able to generate a large set of novel findings, which may serve as experimental targets for biologists and biochemists. Interactive reports are generated to facilitate further exploration of the biological significance of findings. With the high-throughput approach and intgrative tools, the bottlenect for us now shifts to publishing these results. We invite researchers in cancer research community to collaborate with us to publish the findings.

Citation Format: Kaixian Yu, Yun Xu, Ke Tang, Albert Steppi, Jun Zhou, Zheng Ouyang, Jinfeng Zhang. An integrated pipeline for TCGA data analysis. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5275.
Abstract 53: Epigenetic silencing of krebs cycle metabolism in kidney cancer
Authors: Kho Eun-Young , Livi Carolina B. , Buckhaults Phillip , Carobbio Francesca , Nam Hye-Young , Kirkman Richard , Crossman David , Vayalil Praveen K. , Wei Shi , DeBerardinis Ralph J. , Sudarshan Sunil
Published: July 2016

Mounting evidence points to alterations in mitochondrial metabolism in renal cell carcinoma (RCC) including recent large-scale transcriptomic data demonstrating loss of tricarboxylic acid (TCA) cycle enzyme expression with aggressive tumors. Using a metabolomics/transcriptomics approach, we identified loss of mRNA expression of PGC-1α and ERR-γ is associated with tumor progression and worsened outcomes in RCC patients. PGC-1α both induces and cooperates with ERR-γ to promote TCA cycle enzyme expression. Additionally, PGC-1α reconstitution in RCC cells reverses the glycolytic phenotype and suppresses invasive phenotypes. Loss of PGC-1α and ERR-γ is through DNA and histone repressive silencing of the transcription factor PRDM16. Loss of PRDM16 is among the most prominent transcriptional changes in metastatic tumor sites. PRDM16 expression in RCC cells restorers TCA cycle enzyme expression, mitigates the Warburg phenotype, and suppresses in vivo tumorigenesis. Collectively, our data uncover a transcription factor network epigenetically silenced in renal cancer resulting in suppression of mitochondrial metabolism.

Citation Format: Eun-Young Kho, Carolina B. Livi, Phillip Buckhaults, Francesca Carobbio, Hye-Young Nam, Richard Kirkman, David Crossman, Praveen K. Vayalil, Shi Wei, Ralph J. DeBerardinis, Sunil Sudarshan. Epigenetic silencing of krebs cycle metabolism in kidney cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 53.
Abstract 549: Immune profiling uncovers neo-antigens and TCR repertoire responding to WX-mPD-1 and WX-mPD-L1 immune checkpoint inhibitors in syngeneic mouse xenograft models
Authors: Dong Hua , Lu Yan , Zhang Qiyao , Zhang Lingling , Chen Xiaoyue , Yang Guizhu , Yang Shuqun , He Miao , Tang Xuzhen , Qin Yuxin , Zheng Yong , Cai Jiexing , Cang Yong , Yan Shaoyu , Li Jing , Ji Qunsheng
Published: July 2016

Introduction: PD-1 and PD-L1antibodies have shown impressive clinical benefits to cancer patients. However, still a significant number of patients fail to respond to those checkpoint inhibitors. To understand the underlying mechanism, we performed genomic and immune profiling of both tumor tissues and cytotoxic T cells isolated from syngeneic mouse xenograft models treated with in-house WuXi anti-mouse PD-1(WX-mPD-1) and anti-mouse PD-L1 (WX-mPD-L1) antibodies.

Material and methods: Syngeneic mouse xenograft models from melanoma cell lines, CloudmanS91 and B16-F10, were used to evaluate the anticancer efficacy of WX-mPD-1 and WX-mPD-L1 antibodies. Cancer cells and xenograft tumors before and after treatment were collected for RNASeq profiling, from which mRNA expression and DNA mutations were called and neo-antigens were predicted. FACS analysis and TCR sequencing were also performed to examine the tumor infiltrating lymphocytes (TILs) population and TCR repertoire diversity, respectively.

Results: Among forty-eight established syngeneic models, nine had been tested with WX-mPD-1 and WX-mPD-L1 antibodies as single agent or in combination. More importantly, we have investigated the expression, mutation loads, neo-antigens for responsive model from CloudmanS91 and partially responsive model from B16-F10. CloudmanS91 model had 8 fold increases in mutation loads compared with B16-F10 model. Neo-antigens identified in CloudmanS91 cells were further knocked out to dissect their roles in activating immune response to WX-mPD-1 and WX-mPD-L1 antibodies in vivo.

Conclusions: WX-mPD-1 and WX-mPD-L1 antibodies as well as syngeneic tumor models are useful tools for pre-clinical combination and MoA studies for cancer immunotherapy. Analysis of Neo-antigen and TCR repertoire helps us better understand immune response to these antibodies in preclinical settings, and suggest novel approaches to enhance the current immunotherapeutic interventions.

Citation Format: Hua Dong, Yan Lu, Qiyao Zhang, Lingling Zhang, Xiaoyue Chen, Guizhu Yang, Shuqun Yang, Miao He, Xuzhen Tang, Yuxin Qin, Yong Zheng, Jiexing Cai, Yong Cang, Shaoyu Yan, Jing Li, Qunsheng Ji. Immune profiling uncovers neo-antigens and TCR repertoire responding to WX-mPD-1 and WX-mPD-L1 immune checkpoint inhibitors in syngeneic mouse xenograft models. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 549.
Abstract 55: Glucose supports EGFR-mutated lung adenocarcinoma survival by sustaining EGFR stability
Authors: Rho Jin Kyung , Choi Yun Jung , Choi Chang-Min , Lee Jae Cheol
Published: July 2016

Tumor cells are genetically unstable, driven by genetic mutations that stimulate cancer cell proliferation and survival. These genetic mutations are generally linked to particular types of cancer. Oncogenic epidermal growth factor receptor (EGFR) is frequently mutated in non-small cell lung cancer (NSCLC), which is essential for NSCLC development and growth. However, the precise roles of EGFR in lung cancer metabolism are still unclear. Here we show that glucose (Glc) metabolism is critical for EGFR stability required for tumor growth and survival. EGFR mutation leads to a robust increase in Glc uptake, lactate production and glycolytic flux. Moreover, EGFR-mutated NSCLC cell lines are much more sensitive than EGFR WT to inhibition of Glc metabolism through EGFR degradation, suggesting that EGFR mutant NSCLCs rely more heavily than EFGR WT on aerobic glycolysis. Knockdown of EGFR in EGFR-mutated NSCLCs results in a decrease in expression level of genes that encode glycolysis enzymes, indicating that EGFR mutation leads to an increase in Glc metabolism and Glc dependence for growth and survival. Furthermore, inhibition of Glc metabolism shows a significant decrease in intracellular ATP, oxygen consumption and level of TCA cycle intermediates. Indeed, the addition of pyruvate dramatically rescues apoptotic cell death caused by EGFR degradation upon inhibition of Glc metabolism, suggesting that Glc to fuel the TCA cycle is essential for tumor survival. In addition, the inhibition of Glc metabolism overcomes T790M-mediated resistance to EGFR-TKIs. Together, our data suggest that the EGFR mutation mediates the reprogramming of Glc metabolism and this metabolic alteration represents an attractive target for new therapy of EGFR-mutated NSCLC.

Citation Format: Jin Kyung Rho, Yun Jung Choi, Chang-Min Choi, Jae Cheol Lee. Glucose supports EGFR-mutated lung adenocarcinoma survival by sustaining EGFR stability. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 55.
Abstract 559: Small molecule T-reg inhibitors for cancer immunotherapy
Authors: Kumar Suresh , Wu Jian , Wang Liquing , Wang Feng , Kodrasov Matthew P. , Agarwal Saket , Sokirniy Ivan , Yeckley Thomas , Weinstock Joseph , Mattern Michael R. , Hancock Wayne W.
Published: July 2016

Immune evasion is a hallmark feature of tumors as they employ various strategies to suppress the immune system's ability to recognize and destroy cancer cells. T cell checkpoint inhibitors such as anti-PD1 and anti-CTLA-4 antibodies spearhead the immune response against a variety of tumors. Numerous studies suggest that the complex immunosuppressive milieux require the development of additional therapeutic agents to potentiate active drugs and thereby broaden the utility and increase the therapeutic indices of revolutionary immune-oncology treatment modalities. The presence of immunosuppressive regulatory T cells (Tregs) in tumors, keeping tumoricidal Teffector cells in check, signals poor prognosis. Thus, depletion of Tregs or impairment of Treg function is an attractive therapeutic approach for cancer. USP7, a deubiquitylase enzyme implicated as a critical node in several cancer signaling pathways, has recently emerged as an essential factor in maintaining Treg functions. Progenra identified small molecule USP7 inhibitors and employed them to show that Treg specific inhibition of USP7 results in impairment of Treg function leading to immune activation, commensurate with the ablation of Foxp3, a transcription factor that is a target of USP7 and is essential to Treg activation. This USP7 inhibitor class was subsequently lead optimized, and selected USP7 inhibitors were evaluated in ADME/PK studies and shown to impair Treg functions and to exhibit powerful anti-tumor activity against syngeneic lung tumor models in immunocompetent mice. In addition, Progenra's USP7 inhibitors significantly augmented the antitumor activity of anti-PD1 antibody, CAR T cell therapy, and cancer vaccines. These studies provide a strong rationale for the use of USP7 inhibitors in combination therapy protocols to improve the efficacy of currently approved cancer immunotherapy agents.

Citation Format: Suresh Kumar, Jian Wu, Liquing Wang, Feng Wang, Matthew P. Kodrasov, Saket Agarwal, Ivan Sokirniy, Thomas Yeckley, Joseph Weinstock, Michael R. Mattern, Wayne W. Hancock. Small molecule T-reg inhibitors for cancer immunotherapy. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 559.