bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4826: Maternal high butter fat intake heightens mammary cancer risk in offsprings gestationally exposed to bisphenol A at environmentally relevant dose
Authors: Leung Yuet-Kin , Govindarajah Vinothini , Cheong Ana , Song Dan , Zhu Xuegong , Ying Jun , Kendler Ady , Medvedovic Mario , Belcher Scott , Ho Shuk-Mei
Published: July 2017

In utero exposure to bisphenol A (BPA) at 250ug/kg BW was shown to augment mammary cancer risk in rodent models. High fat diet is widely believed to be a major risk of breast cancer (BCa). It is therefore important to investigate whether maternal high fat intake could potentially heighten mammary cancer risk associated to gestational BPA exposure. In this study, we exposed Sprague Dawley rats with different doses of BPA (2.5-2500ug/kg BW) mixed with high butter-based diet (HBF) in the period from preconception to birth. DMBA-PND50 protocol was followed. In the presence of HBF, we observed non-monotonic effects of BPA in cancer incidence as well as in the number of terminal end buds of PND21 mammary glands. Both results were peaked at a dosage of 25ug/kg BW BPA and the effects were significant when compared with HBF alone group. We further analyzed transcriptomic data of microdissected epithelia of PND21 mammary glands from BPA (25ug/kg BW) group as well as HBF group and revealed that two specific cancer networks involving ERK and androgen receptor signaling were dysregulated. DNA methylation is one of the key mechanisms to dysregulate and impair the transcription of two selected top genes, Car7 and Kcnv2, as supported by our bisulfite-sequencing data. These data suggest that in utero BPA exposure could epigenetically modify gene expression and predispose cancer risk during early development. More importantly, some BPA genes are of high human relevance because seven selected BPA genes could dichotomize patients into two groups with significant difference in overall survival in a TCGA cancer cohort. The prognostic power of the genes was further enhanced in the survival analysis of Caucasian with ER positive patients. In conclusion, our data strongly suggest that BPA predisposes higher cancer risk by dysregulating gene expression during early mammary gland development in the presence of HBF, which could facilitate cancer development later in life.

Citation Format: Yuet-Kin Leung, Vinothini Govindarajah, Ana Cheong, Dan Song, Xuegong Zhu, Jun Ying, Ady Kendler, Mario Medvedovic, Scott Belcher, Shuk-Mei Ho. Maternal high butter fat intake heightens mammary cancer risk in offsprings gestationally exposed to bisphenol A at environmentally relevant dose [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4826. doi:10.1158/1538-7445.AM2017-4826
Abstract 4832: Establishment of lung cancer organoid lines as a new preclinical model for lung cancer
Authors: Kim Minsuh , Chun Sung-Min , Mun Hyemin , Suh Young-Ah , Jang Se jin
Published: July 2017

Human cancers are composed of genetically and phenotypically heterogeneous subpopulations organized into a hierarchy of tumor initiating stem cells and various differentiated tumor cell. Cancer organoids seem to be more suitable in vitro models for targeting tumor heterogeneity than cell lines. We established cancer organoid lines from lung cancer patient tissues, recapitulating histology of human cancers in three dimensional culture conditions. Thirteen organoid lines from various lung cancer subtypes including adenocarcinoma, squamous cell carcinoma and small cell carcinoma were compared with their original human tumors in histologic features and genetic profiles. In the IHC staining, it revealed that organoids of adenocarcinomas have typical glandulo-papillary structures or mucin-containing tumor cells. Organoids of squamous cell carcinomas showed keratinization and intercellular bridges. Organoids of small cell carcinoma displayed typical neuroendocrine morphology. Using NGS cancer panel sequencing, the organoid lines maintained genetic characteristics of the original human tumors in major driver genes including EGFR, TP53, and RB mutation. In xenograft experiments, the organoid lines revealed stronger tumorigenesity than direct graft of human tumor tissues. Cancer organoids showed resistance to various anticancer drug treatments. In conclusion, our lung cancer organoid lines are alternative cancer model for preclinical researches recapitulating genotypic and phenotypic heterogeneity of original human tumors.

Note: This abstract was not presented at the meeting.

Citation Format: Minsuh Kim, Sung-Min Chun, Hyemin Mun, Young-Ah Suh, Se jin Jang. Establishment of lung cancer organoid lines as a new preclinical model for lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4832. doi:10.1158/1538-7445.AM2017-4832
Abstract 4841: Suppression of metastasis by KP54 and non-KP54 kisspeptins
Authors: Hampton Kelsey Rebecca , Pounds Keke M. , Trembath Andrew P. , Welch Danny R.
Published: July 2017

KISS1 is a secreted neuropeptide which can act as a metastasis suppressor protein. Expression of KISS1 inhibits metastatic colonization at secondary sites, rendering disseminated cells dormant. In order for KISS1 to suppress metastasis, it must be secreted outside of the cell, where furin cleaves KISS1 into kisspeptins. This cleavage liberates an internal kisspeptin, Kisspeptin-54 (KP54, K67 to F121), which is amidated and has the ability to bind a Gq/11-coupled protein receptor KISS1R. The mechanism of action for KISS1 metastasis suppression has long been assumed to be KP54 interacting with KISS1R. However, expression of KISS1R is not necessary for KISS1 metastasis suppression, and the extracellular processing of KISS1 hints at an alternative hypothesis: a different kisspeptin may be responsible for suppressing metastasis. To test this hypothesis, all possible kisspeptins were generated based on known dibasic cleavage sites (M1-Q145; M1-R56; M1-R67; M1-R124; R56-R66; R67-F121; R56-F121; R56-Q145; R67-Q145; R124-Q145) and were cloned to include the same secretion sequence as nascent KISS1. These kisspeptins (KISS1 Manufactured Peptides, or KMP) were independently expressed in B16-F10 murine melanoma cells. To test for experimental metastasis, B16-F10 cells expressing KMP were injected intravenously into the lateral tail vein of syngeneic mice and the lungs were examined for metastatic load. While KP54 suppressed metastasis, additional KMP lacking the KISS1R binding site (LRF-NH2) were able to completely suppress metastasis (p<0.05). When assessing metastasis-associated traits in vitro, suppressor KMP suppressed motility, increased mitochondrial biomass, and increased oxidative phosphorylation (phenotypes previously reported with KISS1 expression). In particular, one kisspeptin (KMP2, M1 - R56) suppressed metastatic traits in vitro as well as completely suppressing metastasis in vivo. As a whole, these data suggest that metastasis suppression by KISS1 is not necessarily contingent on KISS1R activation, and also supports investigation into additional receptors.

Citation Format: Kelsey Rebecca Hampton, Keke M. Pounds, Andrew P. Trembath, Danny R. Welch. Suppression of metastasis by KP54 and non-KP54 kisspeptins [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4841. doi:10.1158/1538-7445.AM2017-4841
Abstract 486: Production of an unbiased, highly reproducible small RNA library for NGS using a novel circularization technology
Authors: Gonzalez-Plasky Marta , McNulty Amanda , Charizanis Konstantinos , Ning Jing , Jan Edward , Carroll Matthew , Kamberov Emmanuel , Hecker Karl
Published: July 2017

MicroRNA sequencing (miRNA-Seq) is a useful tool for aiding researchers in the examination of miRNA expression patterns, the characterization of novel miRNAs, and for uncovering miRNA-disease associations. Since miRNAs are also unusually well-preserved in a range of samples (e.g. urine, FFPE tissue, plasma), profiling their expression is becoming a powerful diagnostic tool. However, current methods for sequencing miRNA require large amounts of total RNA, are not very reproducible and, more importantly, have considerable systematic bias resulting in loss of many prospective biomarkers. This bias severely affects the trustworthiness of results as libraries are not a true representation of the biological state of the sample.

We have recently developed a technology that efficiently captures miRNA species with extremely low bias. Through a circularization approach, Illumina P5 and P7 adapters are ligated to miRNA, resulting in sequenceable libraries in under 5 hours in a single tube. These libraries are then purified and size-selected prior to sequencing.

Analysis of read distributions of libraries prepared from an equimolar mixture of 963 miRNAs demonstrated the technology is able to efficiently capture all species with high reproducibility between replicates and input amounts (correlation coefficient of ≥0.97). Furthermore, 80-86% of these captured miRNA read numbers fall within a +/- 2-fold variation of the expected read number demonstrating unbiased representation. In contrast, frequency distribution analyses for other technologies reveal that 47-56% of miRNAs are greatly under-represented (i.e. less than 2X fewer reads than expected), 29-38% of miRNAs are represented within a +/- 2-fold variation of the expected read number, and around 15% are over-represented by more than 2X.

These findings highlight the fact that the current technical state of library preparation for miRNA sequencing fails to accurately represent miRNA expression patterns. This in turn leads researchers to perform more costly validation experiments than necessary. Our product is designed and developed for more accurately reflecting the true biological state of a sample. This will provide researchers with a tool that can aid in advancing miRNA sequencing in translational research.

Note: This abstract was not presented at the meeting.

Citation Format: Marta Gonzalez-Plasky, Amanda McNulty, Konstantinos Charizanis, Jing Ning, Edward Jan, Matthew Carroll, Emmanuel Kamberov, Karl Hecker. Production of an unbiased, highly reproducible small RNA library for NGS using a novel circularization technology [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 486. doi:10.1158/1538-7445.AM2017-486
Abstract 4865: The functional and clinicopathological analysis of hypoxia inducible factor-1α (HIF1-α) in head and neck squamous cell carcinoma
Authors: Ikari Yuichi , Ozawa Hiroyuki , Imanishi Yorihisa , Tomita Toshiki , Sekimizu Mariko , Watanabe Yoshihiro , Ito Fumihiro , Saito Shin , Ogawa Kaoru
Published: July 2017

Introduction: Tumor hypoxia is considered to be associated with progression in various cancers. Hypoxia inducible factor-1α (HIF-1α) is one of the molecules which play a key role in tumor hypoxia. High HIF-1α expression is reported to have an impact on poor prognosis and resistance to chemotherapy or radiotherapy in head and neck squamous cell carcinoma (HNSCC). However, the effect and mechanism of HIF-1α on tumor metastasis in HNSCC is still unclear. The aim of the present study was to investigate the molecular targets of HIF1-α relevant to metastasis, and to reveal the impact of HIF1-α on HNSCC metastasis by using clinicopathological assays. Methods: The cell proliferation rate with or without HIF-1 inhibitor (KC7F2) in HNSCC cell lines (FaDu, Detroit512) was evaluated by MTS assay. Gene expression levels related to epithelial-mesenchymal transition (EMT), cancer stem cell (CSC) and HIF-1α in each HNSCC cell lines were measured by quantitative real-time PCR. Immunohistochemical expressions of HIF-1α and clinicopathological factors were analyzed using surgical specimens from 33 patients with hypopharyngeal carcinoma who had undergone trans-oral tumor resection without previous chemotherapy nor radiotherapy.

Results: HIF-1 inhibitor suppressed the cell growth of HNSCC cell lines in a dose dependent manner. HIF-1 inhibitor downregulated the expression of HIF-1α, twist, Oct3/4 and Nanog. In immunohistochemical study, the HIF-1α expression was higher in cancer cells than in normal cells of the adjacent normal mucosa. Univariate analysis revealed that high expression of HIF-1α was significantly correlated with T-classification (P=0.012), lymph node metastasis (P=0.002) and lymphatic invasion (P=0.03).

Conclusion: These results suggest that HIF-1α plays a key role in tumor progression of HNSCC through EMT and CSC properties. From the present study, HIF-1α inhibition may have the possibility to suppress tumor metastasis, making HIF-1α an attractive target for the treatment of HNSCC.

Citation Format: Yuichi Ikari, Hiroyuki Ozawa, Yorihisa Imanishi, Toshiki Tomita, Mariko Sekimizu, Yoshihiro Watanabe, Fumihiro Ito, Shin Saito, Kaoru Ogawa. The functional and clinicopathological analysis of hypoxia inducible factor-1α (HIF1-α) in head and neck squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4865. doi:10.1158/1538-7445.AM2017-4865
Abstract 4880: MAGE gene family expression in pediatric medulloblastoma: frequency and possible therapeutic target
Authors: Collins Rebecca R. , Tacer Klementina Fon , Rakheja Dinesh , Potts Patrick Ryan
Published: July 2017

Background: The melanoma antigen gene family (MAGE) is a group of related genes that were first discovered in malignant melanoma. Among the approximately 40 MAGE genes in the human genome, two thirds are classified as Type I (MAGE-A, MAGE-B, and MAGE-C), referred to also as cancer testis antigens (CTAs), proteins that are restricted to testis but often aberrantly expressed in many adult cancers. Despite their common expression in adult cancers and their oncogenic potential, little is known about their expression and role in pediatric cancers. To determine the frequency of MAGE expression in pediatric medulloblastoma, we measured expression levels of Type I MAGEs in patient tumors. In addition, we examined how MAGE expression in medulloblastoma cells affects their growth and oncogenic potential.

Methods: We searched the Children’s Health pathology database for pediatric medulloblastoma between 2008 - 2015 and were able to recover an adequate amount of RNA from formalin-fixed paraffin-embedded tissue samples of 34 patients. The RNA was converted to cDNA using reverse transcriptase, and the relative expression of 23 Type I MAGE genes was measured by qRT-PCR.

To determine if MAGE expression is critical for medulloblastoma cell survival we knocked down several MAGEs using siRNAs in two MAGE-expressing medulloblastoma cell lines (D283 and DAOY) and evaluated the effect by cell viability (Alamar Blue and Cell titer Glo), apoptosis assays (Annexin V), and colony formation in soft agar.

Results: In 34 medulloblastoma patient samples we found that 10 (29.4%) expressed at least one MAGE gene, and one case had high levels of expression of 10 of the 23 Type I MAGEs. MAGE expression was associated with the non-WNT/SHH (G3/G4) molecular subgroup of tumors in our study. There was no correlation between MAGE expression and histologic subtype. Furthermore, using MAGE-A/-B2 positive medulloblastoma cell lines, we showed that siRNA knock down of multiple MAGE-As and -B2 decreased cell viability by 15 - 80%, increased apoptosis (-B2), and decreased growth of cells in soft agar.

Conclusion: Type I MAGE genes are expressed in 30% of pediatric medulloblastoma in our study. In MAGE positive medulloblastoma cells, MAGE genes are important for cell survival and clonogenic growth. The decreased cell survival after MAGE knockdown appears to be related, at least in part, to increased apoptosis. Our results indicate that targeting of MAGEs in medulloblastoma may be a potential therapeutic option.

Citation Format: Rebecca R. Collins, Klementina Fon Tacer, Dinesh Rakheja, Patrick Ryan Potts. MAGE gene family expression in pediatric medulloblastoma: frequency and possible therapeutic target [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4880. doi:10.1158/1538-7445.AM2017-4880
Abstract 4926: Gut microbiota regulates cisplatin mediated cachexia and systemic toxicity
Authors: Roy Soumen , Neves Rodrigo Das , Dzutsev Amiran , Smith Carolyne , Edwards Bathai , Dawson Miranda , Difilippantonio Simone , Smith Loretta , Huang April , Kim Young , Trinchieri Giorgio
Published: July 2017

Chemotherapy induced toxicity severely affect the cancer survivors and lowers the quality of life. By 2020, there will be more than 18 million of cancer survivors all over the world. Majority of them might develop long term nephrotoxicity, ototoxicity and gut toxicity. In addition, chemotherapy may also facilitate the initiation and progression of cachexia. Recent studies have shown that gut microbiota modulates the efficacy of anti-cancer chemotherapy, however very limited knowledge is available regarding the role of gut microbiota in regulating systemic toxicity and cachexia. We hypothesized that gut microbiota modulates cisplatin induced systemic toxicity as well as cachexia. Four groups (n=10 in each group) of 6-8 weeks old C57B/6 mice were treated with cisplatin, cisplatin+antibiotics cocktails (ABX), ABX only and control. ABX cocktail contained primaxin, vancomycin and neomycin. This experiment was validated using C57B/6 germ free mice. We performed anti-p-γ-H2AX based toxicity assay for DNA damage. In addition, we performed immunohistochemistry for studying cachexia. H&E staining and DNA damage were assessed by light and super resolution confocal microscopy (Zeiss 880) and quantified by 3-D reconstruction using IMARIS. We also performed high content imaging to evaluate DNA damage in bone marrow and spleen. Body weight and kidney blood content were also analyzed to determine the degree of toxicity. Depletion of gut microbiota significantly reduced nephrotoxicity and gut toxicity. In addition, we found that depletion of gut microbiota prevents muscle and adipose tissue loss by down-regulating UCP-1and PGC-1α in adipose tissue and MURF-1 and Atrogin-1 in muscle. Experiments are confirmed by using Germ free mice. Our data suggest that modulation of gut microbiota may be utilized to reduce chemotherapy associated cachexia and systemic toxicity.

Citation Format: Soumen Roy, Rodrigo Das Neves, Amiran Dzutsev, Carolyne Smith, Bathai Edwards, Miranda Dawson, Simone Difilippantonio, Loretta Smith, April Huang, Young Kim, Giorgio Trinchieri. Gut microbiota regulates cisplatin mediated cachexia and systemic toxicity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4926. doi:10.1158/1538-7445.AM2017-4926
Abstract 493: Pathologic findings at RRSO in germline BRCA mutation carriers with breast cancer: significance of bilateral RRSO at the optimal age in germline BRCA mutation carriers
Authors: Lee YoungJae , Kim YongMan , Lee ShinWha
Published: July 2017

Objective: Most BRCA1/2 carriers do not undergo risk-reducing salpingo-oophorectomy (RRSO) by the recommended age. This study aimed to find the incidence of precursor lesions and cancer after RRSO.

Methods: We retrospectively reviewed breast cancer patients identified as BRCA mutation carriers who underwent RRSO at Asan Medical Center, Seoul, Korea, from 2010 to 2014. From 2013, all cases were examined according to the SEE/FIM protocol and underwent immunohistochemically (IHC) staining. RRSO was performed in 63 patients, 27 in 2010-2012 and 36 in 2013-2014.

Results: The median age at RRSO was 46.5 years (32-73 years). Occult invasive cancer was detected in 8 patients, of ovarian origin in 5 and of tubal origin in 3. All occult invasive cancer cases with metastasis were detected in patients older than 40 years. Of the 36 patients from the 2013-2014 cohort, 7 showed p53 overexpression, 1 showed Ki-67 overexpression, 2 showed serous tubal intraepithelial carcinoma (STIC), and 3 showed occult cancer. The detection rate of precursor lesions or cancer was 36.1% (13/36). In the analysis according to age, precursor lesions were more common in BRCA1 mutation carriers younger than 40 years old (66.7% vs 20.0%). In BRCA2 mutation carriers, precursor lesions were only detected in those older than 40 years of age, indicating the possible faster occurrence of precursor lesions in BRCA1 mutation carriers.

Conclusions: Many patients still tend to delay RRSO until after they are 40 years old. Our findings support the significance of RRSO before the age of 40 in germline BRCA mutation carriers.

Citation Format: YoungJae Lee, YongMan Kim, ShinWha Lee. Pathologic findings at RRSO in germline BRCA mutation carriers with breast cancer: significance of bilateral RRSO at the optimal age in germline BRCA mutation carriers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 493. doi:10.1158/1538-7445.AM2017-493
Abstract 4941: Lysophosphatidic acid receptor signaling and pancreatic adenocarcinoma tumor microenvironment
Authors: Skill Nicholas J. , Maluccio Mary A.
Published: July 2017

Introduction: Changes in lipid biosynthesis has attracted significant attention in the study of cancer. Its value in our understanding of pancreatic cancer has not yet been realized. This study examined lysophosphatidic acid (LPA) variant biosynthesis in patients with stage II-III pancreatic ductal adenocarcinoma (PDAC), evaluating serum, bile, and tissue levels of LPA, its receptors, and downstream metabolites. LPA, a product of phospholipase D activity of the enzyme autotaxin (ATX) is a potent mitogenic agent. Data suggests that changes in lipid biosynthesis contribute to the tumor microenvironment in PDAC.

Methods: PDAC, matched adjacent non-malignant tissues, gall bladder bile and serum samples were collected from patients with Stage II-III pancreatic cancer at the time of surgery. Control bile and serum were collected from patients undergoing cholecystectomy for benign disease and living renal donors respectively.

Results: Serum and biliary LPA levels were increased in patients with PDAC when compared to controls. ATX activity, and LPA receptors, LPAR1, LPAR2, and LPAR5 were greater in PDAC adjacent pancreatic tissue when compared to PDAC tissue. In contrast, TNFα receptors TNFα1 and TNFα2 and LPA receptor LPAR3 receptor expression was greater in PDAC lesions when compared to adjacent tissue.

Conclusion: PDAC patients display a modified LPA biosynthesis profile, when compared to non-PDAC patients, potentially providing a paracrine-signaling cascade to promote pancreatic tumorigenesis through increased expression of mitogenic cytokines and growth factors.

Citation Format: Nicholas J. Skill, Mary A. Maluccio. Lysophosphatidic acid receptor signaling and pancreatic adenocarcinoma tumor microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4941. doi:10.1158/1538-7445.AM2017-4941
Abstract 496: Ubiquitin specific peptidases 37 promotes constitutive replication fork movement by stabilizing Chk1 via its deubiquitination
Authors: Singh Mayank , Burrows Amy C. , Dickson Andrew , Komal Komal , Pal Debjani , Summers Matthew K.
Published: July 2017

Protein deubiquitination controls many intracellular processes, including cell cycle progression, transcriptional activation, and signal transduction. Ubiquitin specific peptidases (USPs) remove ubiqiuitin tags from target proteins to control both protein fate and function. USP37 (Ubiquitin specific peptidase 37) has been implicated in cancer and we have previously shown that antagonizes the tumor suppressor APCCDH1and promotes S phase entry. However, the role of USP37 during S-phase was unknown. Here, we report that in cells experiencing replication stress USP37 overexpression confers survival advantage while its depletion enhances sensitivity. USP37 overexpressing cells were able to resolve different DNA damage markers much more effectively then the control cells or cells in which USP37 was depleted. Mechanistically, our data indicate that USP37 binds and stabilizes the active form of CHK1 and deubiquitinates Chk1 to increase its stability and promote the checkpoint response. USP37 overexpression results in constitutive replication fork movement and long tract DNA synthesis while USP37 depleted cells were unable to carry out long tract DNA synthesis. Notably, expression of low level of Chk1 in USP37 depleted cells rescues cell survival and DNA damage response. Overall our data suggest that inhibition of USP37 may represent a novel mechanism to modulate Chk1 activity.

Note: This abstract was not presented at the meeting.

Citation Format: Mayank Singh, Amy C. Burrows, Andrew Dickson, Komal Komal, Debjani Pal, Matthew K. Summers. Ubiquitin specific peptidases 37 promotes constitutive replication fork movement by stabilizing Chk1 via its deubiquitination [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 496. doi:10.1158/1538-7445.AM2017-496
Abstract 4967: RAF kinase inhibition synergizes with MEK inhibitors in KRAS mutant tumors
Authors: Yen Ivana , Klijn Christiaan , Shanahan Frances , Merchant Mark , Orr Christine , Hunsaker Thomas , Durk Matthew , La Hank , Zhang Xiaolin , Martin Scott , Lin Eva , Chan John , Yu Yihong , Gustafson Amy , Rudolph Joachim , Malek Shiva
Published: July 2017

While activating mutations in the KRAS oncogene frequently drive tumorigenesis in human cancers (40% CRC, 20% NSCLC) through constitutive activation of the MAPK pathway, there are currently no targeted treatments available for KRAS mutant cancers. Inhibitors of the individual nodes of the MAPK pathway have been developed, but these molecules have been largely ineffective against KRAS mutant tumors in the clinic. Multiple studies have shown rational combinations of MAPK inhibitors may have anti-tumor activity in KRAS mutant models. In order to understand the versatility of combining RAF inhibitors in this context, we conducted a library screen consisting of 430 small molecule tool compounds in combination with RAF inhibitor AZ-628. Here we show: RAF inhibitors combine especially well with other MAPK pathway inhibitors in KRAS mutant tumor cells. In particular, Type II RAF inhibitors are synergistic with the MEK inhibitor Cobimetinib in vitro and exhibit tumor regressing efficacy in xenograft mouse model studies in vivo. Mechanistically, we have found the MEK inhibitor disables ERK induced negative feedback on the MAPK pathway resulting in activation of CRAF in a KRAS dependent manner. The combination of RAF with MEK inhibition blunts KRAS-dependent activation of CRAF kinase activity and robustly inhibits MAPK signaling, thereby driving efficacy in KRAS mutant tumors. Broad cell line profiling with the combination of RAF and MEK inhibitors demonstrates that a majority of KRAS mutant lung and colorectal tumor lines exhibit synergy with the combination. Therefore, combining a Type II pan-RAF inhibitor with a MEK inhibitor has the potential to improve the therapeutic outcome in KRAS mutant cancers.

Citation Format: Ivana Yen, Christiaan Klijn, Frances Shanahan, Mark Merchant, Christine Orr, Thomas Hunsaker, Matthew Durk, Hank La, Xiaolin Zhang, Scott Martin, Eva Lin, John Chan, Yihong Yu, Amy Gustafson, Joachim Rudolph, Shiva Malek. RAF kinase inhibition synergizes with MEK inhibitors in KRAS mutant tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4967. doi:10.1158/1538-7445.AM2017-4967
Abstract 4970: Oxidative phosphorylation as a target in triple negative breast cancer therapy
Authors: Meric-Bernstam Funda , Evans Kurt , Zheng Xiaofeng , Su Xiaoping , Yuca Erkan , Scott Stephen , Akcakanat Argun , Ueno Naoto , Lim Bora , Litton Jennifer , Valero Vicente , Symmans Fraser , Hortobagyi Gabriel , Perou Charles , Tripathy Debu , Draetta Guilio , Marszalek Joe , Gonzalez-Angulo Ana Maria , Moulder Stacy
Published: July 2017

Altered cellular metabolism is a hallmark of cancer. It is increasingly recognized that selected tumors are dependent on oxidative phosphorylation (OXPHOS). However, the role of OXPHOS in TNBC is not well understood. We performed RNA sequencing in pre-treatment biopsies from 43 patients with operable triple negative breast cancer (TNBC) who received sequential taxane- and anthracycline-based neoadjuvant chemotherapy. At a median follow-up of 63 months, 14 patients recurred and 12 patients died. At a false discovery rate of 0.05, 33 genes were differentially expressed between the patients who did and did not have a subsequent recurrence. Ingenuity pathway analysis demonstrated that one of the top canonical pathways that differed was higher expression of oxidative phosphorylation signature (p=5.89E-0.7). The patients who recurred had significantly higher levels of mitochondrial genes: MT-ND1 (adjusted p or FDR-BH; q=0.007); MT-ND5 (q=0.03) and MT-ND4 (q=0.04). Further, 21 genes were differentially expressed between patients based on survival, including MT-ND5 (q=0.001); MT-ND4 (q=0.005), MT-ND4L (q=0.015), MT-ND6 (q=0.018), and MT-ATP6 (P=0.03). Top canonical pathway that was differentially expressed based on survival was oxidative phosphorylation (p=9.98E-10). We therefore sought to determine the efficacy of IACS-10759, a novel inhibitor of OXPHOS, in 10 different TNBC patient-derived xenografts representing different gene expression based Lehmann TNBC subtypes. Growth inhibition was observed in multiple subtypes, with regression in one basal-like 1(BL1) 1 model, and stabilization of growth in multiple BL1 and immunomodulatory expression subtypes. Taken together, our data suggests that high OXPHOS is associated with higher recurrence and lower survival. OXPHOS is a promising target in several TNBC subtypes. A Phase I trial of IACS-10759, a potent inhibitor of complex I of oxidative phosphorylation, in leukemia is ongoing and planned in TNBC and other solid tumors.

Citation Format: Funda Meric-Bernstam, Kurt Evans, Xiaofeng Zheng, Xiaoping Su, Erkan Yuca, Stephen Scott, Argun Akcakanat, Naoto Ueno, Bora Lim, Jennifer Litton, Vicente Valero, Fraser Symmans, Gabriel Hortobagyi, Charles Perou, Debu Tripathy, Guilio Draetta, Joe Marszalek, Ana Maria Gonzalez-Angulo, Stacy Moulder. Oxidative phosphorylation as a target in triple negative breast cancer therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4970. doi:10.1158/1538-7445.AM2017-4970
Abstract 4981: Extracellular spacer and co-stimulatory domains define target sensitivity and persistence of CAR T cells for the treatment of PSCA+ bone metastatic prostate cancer
Authors: Kennewick Kelly , Tilakawardane Dileshni , Gerdts Ethan , Murad John , Park Anthony , Jeang Brook , Yamaguchi Yukiko , Forman Stephen J. , Priceman Saul
Published: July 2017

Advancing chimeric antigen receptor (CAR)-engineered adoptive T cells for the treatment of solid cancers is a major focus in the field of immunotherapy, given impressive recent clinical responses in hematological malignancies. Prostate cancer should be amenable to CAR-based immunotherapy given that several tumor antigens, including prostate stem-cell antigen (PSCA), are widely over-expressed in metastatic disease. The selectivity of CARs for solid cancers is crucial, however, due to the absence of truly restricted tumor antigen expression and potential safety concerns with “on-target off-tumor” activity. Here, we show that optimized co-stimulatory signaling and extracellular spacer domains are essential in defining a CAR’s selectivity towards tumor cells that over-express the target antigen. The 4-1BB co-stimulatory domain in PSCA-CARs confers enhanced tumor selectivity with dampened yet dose-responsive cytokine production, reduced T cell exhaustion phenotype, and equivalent tumor killing ability compared to PSCA-CARs containing the CD28 co-stimulatory domain. CARs containing a short extracellular spacer demonstrate the most selective killing of high PSCA-expressing tumors but are unable to produce the inflammatory cytokines important for a complete anti-tumor response. We show that longer extracellular spacers in PSCA-CARs are necessary for both tumor killing and cytokine production. PSCA-CARs exhibit robust in vivo anti-tumor activity in subcutaneous and orthotopic bone-metastatic patient-derived prostate cancer xenograft models, and 4-1BB-containing CARs show superior persistence in controlling metastatic disease compared with CD28-containing CARs. Our study demonstrates the critical impact of CAR components in defining an optimized tumor-selective CAR T cell, and also highlights the importance of clinically relevant animal models in developing effective solid cancer CAR T cell therapies.

Citation Format: Kelly Kennewick, Dileshni Tilakawardane, Ethan Gerdts, John Murad, Anthony Park, Brook Jeang, Yukiko Yamaguchi, Stephen J. Forman, Saul Priceman. Extracellular spacer and co-stimulatory domains define target sensitivity and persistence of CAR T cells for the treatment of PSCA+ bone metastatic prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4981. doi:10.1158/1538-7445.AM2017-4981
Abstract 5001: Functional biomarker discovery from cancer patient serum with PEP technology
Authors: Wang David L. , Fu Guofeng , Davies Micheal , Wang Xing
Published: July 2017

Proteins play essential roles in numerous biological processes and being able to pinpoint functional differences between cell or tissue samples can greatly aid in understanding disease processes and metabolic changes. The PEP technology allows systematic analysis of protein functions within a proteome. Hundreds of functional proteins can be separated and functionally assayed to generate a comprehensive three-dimensional landscape of protein families such as protein kinases, phosphatases, proteinases and oxido-reductases. This information can then be integrated into other genomic and proteomic knowledge bases to provide further insight of important biological processes such as cancer development and aging.

In the PEP technology, complex protein mixtures are first separated by a modified two-dimensional gel electrophoresis process, giving enhanced resolution while still maintaining protein function. This is followed by an efficient protein transfer to a specially designed 1536-well Protein Elution Plate. After further transfer of the samples from the PEP plate to multiple 384-well microplates, functional assays can be performed on each well to generate an enzyme activity profile displayed in 3-D. Protein components of each well can be further characterized by mass spectrometry if desired.

Using the PEP technology, functional biomarker candidates have been identified from lung and breast cancer patient serum. Both qualitative and quantitative differences of metabolic enzymes and proteases were observed when comparing the cancer patient serum and normal serum. Some of the active enzymes were identified by mass spectrometry and validated in selected bioassays. It is believed that this functional proteomics technology provides a unique approach in the discovery of potential cancer biomarkers for diagnostic and prognostic applications.

Citation Format: David L. Wang, Guofeng Fu, Micheal Davies, Xing Wang. Functional biomarker discovery from cancer patient serum with PEP technology [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5001. doi:10.1158/1538-7445.AM2017-5001
Abstract 5008: The brain microenvironment mediates resistance in luminal breast cancer to PI3K inhibition through HER3 activation
Authors: Ferraro Gino B. , Kodack David P. , Askoxylakis Vasileios , Sheng Qing , Badeaux Mark , Goel Shom , Qi Xiaolong , Shankaraiah Ram , Cao Alexander Z. , Ramjiawan Rakesh R. , Bezwada Divya , Patel Bhushankumar , Song Youngchul , Costa Carlotta , Naxerova Kamila , Wong Christina , Kloepper Jonas , Das Rita , Tam Angela , Tanboon Jantima , Duda Dan G. , Miller Ryan C. , Siegel Marni B. , Anders Carey K. , Sanders Melinda , Estrada Valeria M. , Schlegel Robert , Arteaga Carlos L. , Brachtel Elena , Huang Alan , Fukumura Dai , Engelman Jeffrey A. , Jain Rakesh K.
Published: July 2017

Brain metastases represent a devastating progression of luminal breast cancer. While targeted therapies are often effective systemically, they fail to adequately control brain metastases. In preclinical models that faithfully recapitulate the disparate clinical responses in these microenvironments, we observed that brain metastases evade PI3K inhibition despite efficient drug delivery. In comparison to extracranial disease, there is increased HER3 expression and phosphorylation in the brain lesions. HER3 blockade overcomes the resistance of both HER2-amplified and/or PIK3CA-mutant breast cancer brain metastases to PI3K inhibitors, leading to striking tumor growth delay and significant improvement of mouse survival. Collectively, these data provide a mechanistic basis underlying therapeutic resistance in the brain microenvironment and identify rapidly translatable treatment strategiesfor HER2-amplified and/or PIK3CA-mutant breast cancer brain metastases.

Citation Format: Gino B. Ferraro, David P. Kodack, Vasileios Askoxylakis, Qing Sheng, Mark Badeaux, Shom Goel, Xiaolong Qi, Ram Shankaraiah, Alexander Z. Cao, Rakesh R. Ramjiawan, Divya Bezwada, Bhushankumar Patel, Youngchul Song, Carlotta Costa, Kamila Naxerova, Christina Wong, Jonas Kloepper, Rita Das, Angela Tam, Jantima Tanboon, Dan G. Duda, Ryan C. Miller, Marni B. Siegel, Carey K. Anders, Melinda Sanders, Valeria M. Estrada, Robert Schlegel, Carlos L. Arteaga, Elena Brachtel, Alan Huang, Dai Fukumura, Jeffrey A. Engelman, Rakesh K. Jain. The brain microenvironment mediates resistance in luminal breast cancer to PI3K inhibition through HER3 activation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5008. doi:10.1158/1538-7445.AM2017-5008
Abstract 5013: A 3D culture system identifies a new mode of cetuximab resistance and disease-relevant genes in colorectal cancer
Authors: Singh Bhuminder , Li Cunxi , Graves-Deal Ramona , Ma Haiting , Starchenko Alina , Fry William H. , Lu Yuanyuan , Wang Yang , Bogatcheva Galina , Khan Mohseen P. , Milne Ginger L. , Zhao Shilin , Ayers Gregory D. , Li Nenggan , Washington Mary K. , Yeatman Timothy J. , McDonald Oliver G. , Liu Qi , Coffey Robert J.
Published: July 2017

We previously reported that single cells from a human colorectal cancer (CRC) cell line (HCA-7) formed either hollow single-layered polarized cysts or solid spiky masses when plated within type-I collagen in 3D. To begin in-depth analyses into whether clonal cysts and spiky masses possessed divergent malignant properties, individual colonies of each morphology were isolated and expanded. The lines thus derived faithfully retained the parental cystic and spiky morphologies and were termed CC (cystic) and SC (spiky), respectively. Although both CC and SC expressed EGF receptor (EGFR), cetuximab strongly inhibited growth of CC, whereas SC was resistant to growth inhibition and this was coupled to increased tyrosine phosphorylation of MET and RON. Addition of the dual MET/RON tyrosine kinase inhibitor, crizotinib, to cetuximab in SC restored cetuximab sensitivity. To characterize genome-wide divergence between CC and SC, we performed comprehensive genomic and transcriptomic analysis of CC and SC in 3D. One of the most upregulated genes in CC was the tumor suppressor 15-PGDH/HPGD and the most upregulated gene in SC was versican (VCAN) in 3D and xenografts. Analysis of a human CRC tissue microarray showed that epithelial, but not stromal, VCAN staining strongly correlated with reduced survival, and combined epithelial VCAN and absent HPGD staining portended a poorer prognosis. Thus, with this 3D system, we have identified a new mode of cetuximab resistance and a potential prognostic marker in CRC. As such, this represents a potentially powerful system to identify additional therapeutic sensitivities and disease-relevant genes for CRC.

Citation Format: Bhuminder Singh, Cunxi Li, Ramona Graves-Deal, Haiting Ma, Alina Starchenko, William H. Fry, Yuanyuan Lu, Yang Wang, Galina Bogatcheva, Mohseen P. Khan, Ginger L. Milne, Shilin Zhao, Gregory D. Ayers, Nenggan Li, Mary K. Washington, Timothy J. Yeatman, Oliver G. McDonald, Qi Liu, Robert J. Coffey. A 3D culture system identifies a new mode of cetuximab resistance and disease-relevant genes in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5013. doi:10.1158/1538-7445.AM2017-5013
Abstract 5018: Dissecting antigen-specific T-cell response in a novel mouse model of colon cancer
Authors: Beytagh Mary C. , Westcott Peter M. , Smith Olivia C. , Roper Jatin , Yilmaz Omer , Jacks Tyler E.
Published: July 2017

Immune checkpoint blockade therapies, which unleash the immune system’s ability to recognize and eliminate cancer cells, are currently among the most promising treatments for cancer. Antibodies that block the immune inhibitory receptors CTLA4 and PD1 have shown clinical success, with objective response rates over 20% and some durable responses exceeding 10 years. However, some cancers, such as colorectal cancer (CRC), have proven particularly refractory to immunotherapy. Although it is appreciated that the colon is an immunosuppressive environment, the mechanism and extent of immune suppression remains unclear. Studies of human CRC tumors have shown that T cell infiltration is an important predictor of clinical outcome, yet current mouse models of CRC are not particularly well suited to the mechanistic study of T cell responses against tumor-specific antigens. Syngeneic cell line transplants, by far the most widely used models to study adaptive immune response in CRC, are characterized by a highly complex tumor antigen landscape, and have not employed orthotopic transplant into the colon. In order to track highly specific T cell responses in CRC, we have employed a novel colon organoid-based system, in which transformed organoids expressing model antigens are orthotopically transplanted into the wall of the colon via colonscopy-guided injection, a method pioneered in the lab of Omer Yilmaz. Organoids were harvested from VillinCreER; Apcfl/fl mice and grown in the presence of Wnt ligand; addition of tamoxifen and removal of Wnt from the media facilitated expression of the colon-specifc Cre, deletion of Apc, and selection of transformed organoids capable of growing in the absence of Wnt. To induce expression of model antigens, organoids were infected with a lentiviral vector that, in addition to the fluorescent reporter FusionRed, expressed the T cell antigens SIYRYYGL and two antigens from ovalbumin - SIINFEKL and OVA323-339 - fused to the C-terminus of luciferase (FRLucOS), enabling us to monitor antigen expression in vivo. Organoids were then sorted and injected into the wall of the colon. Previous work by Michael DuPage, in which he expressed the LucOS construct in an autochthonous lung cancer and sarcoma mouse model, provided key insights into the dynamic nature of T cell response against cancer cells. The novel model system described here provides a powerful new tool to dissect antitumor T cell responses in the poorly understood context of CRC.

Citation Format: Mary C. Beytagh, Peter M. Westcott, Olivia C. Smith, Jatin Roper, Omer Yilmaz, Tyler E. Jacks. Dissecting antigen-specific T-cell response in a novel mouse model of colon cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5018. doi:10.1158/1538-7445.AM2017-5018
Abstract 5029: Pharmacogenetics of doxorubicin, gemcitabine and docetaxel in the GeDDiS soft tissue sarcoma trial
Authors: Jamieson David , Seddon Beatrice , Kuever Katja , Dehbi Hakim-Moulay , Strauss Sandra , Beare Sandy , Veal Gareth
Published: July 2017

Introduction

The treatment of locally advanced or metastatic soft tissue sarcoma remains a significant clinical challenge, with overall survival rates of approximately 12 months observed with current first line palliative chemotherapy. A recently completed randomised phase III clinical trial entitled GeDDiS (ISRCTN07742377), was designed to compare first line treatment with gemcitabine in combination with docetaxel (GemDoc) versus current standard treatment with doxorubicin. A pharmacogenetic sub-study was incorporated into the trial to assess potential impact of single nucleotide polymorphisms (SNPs) in genes associated with the pharmacology of the three drugs.

Methods

A total of 240 patients were recruited to the sub-study with 119 on the doxorubicin arm and 121 on the GemDoc arm. A 4 ml blood sample was taken from each participant and genomic DNA extracted. Individual candidate SNPs were assessed by Taqman PCR in 7 genes associated with doxorubicin pharmacology, 7 genes associated with gemcitabine and 5 genes associated with the pharmacology of docetaxel. Association between the SNPs and efficacy and toxicity was assessed.

Results

SNPs within the solute transporter SLC22A16, associated with the intracellular influx of doxorubicin, were associated with worse PFS (HR = 1.72, p=0.04) and decreased frequency of grade 3/4 adverse events (71% vs 48%, p=0.04) in the doxorubicin arm of the study but not in the GemDoc arm.

Conclusions

The association of genetic variants of SLC22A16 with decreased efficacy and decreased incidence of toxicity is consistent with a loss of function in the transporter and previous observation of increased AUC in pharmacokinetic studies assuming that distribution of the drug into target and collateral tissues is restricted.

Citation Format: David Jamieson, Beatrice Seddon, Katja Kuever, Hakim-Moulay Dehbi, Sandra Strauss, Sandy Beare, Gareth Veal. Pharmacogenetics of doxorubicin, gemcitabine and docetaxel in the GeDDiS soft tissue sarcoma trial [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5029. doi:10.1158/1538-7445.AM2017-5029