bluuebunny/crossref_metadata_2025 huggingface dataset:
Supplemental Tables 1-3 from CDK4/6 Inhibitors Sensitize Rb-positive Sarcoma Cells to Wee1 Kinase Inhibition through Reversible Cell-Cycle Arrest
Authors: Francis Ashleigh M. , Alexander Angela , Liu Yanna , Vijayaraghavan Smruthi , Low Kwang Hui , Yang Dong , Bui Tuyen , Somaiah Neeta , Ravi Vinod , Keyomarsi Khandan , Hunt Kelly K.
Published: April 2023

Supplemental Table 1 - Cell line culture conditions and HTSA plating density Supplemental Table 2 - Concentrations of drugs used in HTSA assays Supplemental Table 3 - PDX information
Figure S4 from Acquired Resistance Mechanisms to Combination Met-TKI/EGFR-TKI Exposure in Met-Amplified EGFR-TKI–Resistant Lung Adenocarcinoma Harboring an Activating EGFR Mutation
Authors: Yamaoka Toshimitsu , Ohmori Tohru , Ohba Motoi , Arata Satoru , Kishino Yasunari , Murata Yasunori , Kusumoto Sojiro , Ishida Hiroo , Shirai Takao , Hirose Takashi , Ohnishi Tsukasa , Sasaki Yasutsuna
Published: April 2023

Although there were no additional mutations, additional Met gene amplification was determined in DR6 cells compared to MET1000 cells.
Supplementary Figure Legends from Gastric Adenocarcinomas Express the Glycosphingolipid Gb3/CD77: Targeting of Gastric Cancer Cells with Shiga Toxin B-Subunit
Authors: Geyer Philipp Emanuel , Maak Matthias , Nitsche Ulrich , Perl Markus , Novotny Alexander , Slotta-Huspenina Julia , Dransart Estelle , Holtorf Anne , Johannes Ludger , Janssen Klaus-Peter
Published: April 2023

Legends for supplementary figures
Supplementary Figure S2 from Interference with the HSF1/HSP70/BAG3 Pathway Primes Glioma Cells to Matrix Detachment and BH3 Mimetic–Induced Apoptosis
Authors: Antonietti Patrick , Linder Benedikt , Hehlgans Stephanie , Mildenberger Iris C. , Burger Michael C. , Fulda Simone , Steinbach Joachim P. , Gessler Florian , Rödel Franz , Mittelbronn Michel , Kögel Donat
Published: April 2023

Enhanced cytochrome C release in U251 BAG3KD cells.
Supplementary Figure S8 from Metformin Synergizes with BCL-XL/BCL-2 Inhibitor ABT-263 to Induce Apoptosis Specifically in p53-Defective Cancer Cells
Authors: Li Xinzhe , Li Bo , Ni Zhenhong , Zhou Peng , Wang Bin , He Jintao , Xiong Haojun , Yang Fan , Wu Yaran , Lyu Xilin , Zhang Yan , Zeng Yijun , Lian Jiqin , He Fengtian
Published: April 2023

Cotreatment with metformin and ABT-263 more remarkably elevates SASP components and p-ATM in p53-WT cancer cells compared to p53-defective cancer cells and the cotreatment increases p53 in p53-WT cancer cells.
Supplemental Figure 1 from Expression of GRP78, Master Regulator of the Unfolded Protein Response, Increases Chemoresistance in Pancreatic Ductal Adenocarcinoma
Authors: Gifford Jenifer B. , Huang Wei , Zeleniak Ann E. , Hindoyan Antreas , Wu Hong , Donahue Timothy R. , Hill Reginald
Published: April 2023

GRP78 induction following Gemcitabine treatment.
Supplementary Figure S7 from Metformin Synergizes with BCL-XL/BCL-2 Inhibitor ABT-263 to Induce Apoptosis Specifically in p53-Defective Cancer Cells
Authors: Li Xinzhe , Li Bo , Ni Zhenhong , Zhou Peng , Wang Bin , He Jintao , Xiong Haojun , Yang Fan , Wu Yaran , Lyu Xilin , Zhang Yan , Zeng Yijun , Lian Jiqin , He Fengtian
Published: April 2023

Metformin and ABT-263 synergistically inhibit the growth of p53-defective (but not p53-WT) cancer cells and the metformin/ABT-263 combination more effectively elicits apoptosis in p53-defective xenograft tumors compared to p53-WT xenograft tumors.
Supplementary Figure S4 from Metformin Synergizes with BCL-XL/BCL-2 Inhibitor ABT-263 to Induce Apoptosis Specifically in p53-Defective Cancer Cells
Authors: Li Xinzhe , Li Bo , Ni Zhenhong , Zhou Peng , Wang Bin , He Jintao , Xiong Haojun , Yang Fan , Wu Yaran , Lyu Xilin , Zhang Yan , Zeng Yijun , Lian Jiqin , He Fengtian
Published: April 2023

Input analysis of corresponding Co-IP.
Figure S1 from Vessel-Targeted Chemophototherapy with Cationic Porphyrin-Phospholipid Liposomes
Authors: Luo Dandan , Geng Jumin , Li Nasi , Carter Kevin A. , Shao Shuai , Atilla-Gokcumen G. Ekin , Lovell Jonathan F.
Published: April 2023

Possible structures of oxidized DOTAP matching molecular weights following phototreatment, observed by HPLC-MS.
Complete Supporting Data from Vessel-Targeted Chemophototherapy with Cationic Porphyrin-Phospholipid Liposomes
Authors: Luo Dandan , Geng Jumin , Li Nasi , Carter Kevin A. , Shao Shuai , Atilla-Gokcumen G. Ekin , Lovell Jonathan F.
Published: April 2023

Complete Supporting Data
Table S2 from Insulin-Mediated Signaling Facilitates Resistance to PDGFR Inhibition in Proneural hPDGFB-Driven Gliomas
Authors: Almiron Bonnin Damian A. , Ran Cong , Havrda Matthew C. , Liu Huan , Hitoshi Yasuyuki , Zhang Zhonghua , Cheng Chao , Ung Matthew , Israel Mark A.
Published: April 2023

Forward and reverse primers sequences.
Figure S1 from Insulin-Mediated Signaling Facilitates Resistance to PDGFR Inhibition in Proneural hPDGFB-Driven Gliomas
Authors: Almiron Bonnin Damian A. , Ran Cong , Havrda Matthew C. , Liu Huan , Hitoshi Yasuyuki , Zhang Zhonghua , Cheng Chao , Ung Matthew , Israel Mark A.
Published: April 2023

Developing a proneural glioma mouse model of resistance to targeted therapy using the established doxycycline regulatable hPDGFB transgenic mouse model of glioma, GFAP/tTA:TRE/hPDGFB.
Supplementary Figure S4 from Afatinib against Esophageal or Head-and-Neck Squamous Cell Carcinoma: Significance of Activating Oncogenic HER4 Mutations in HNSCC
Authors: Nakamura Yu , Togashi Yosuke , Nakahara Hirokazu , Tomida Shuta , Banno Eri , Terashima Masato , Hayashi Hidetoshi , de Velasco Marco A. , Sakai Kazuko , Fujita Yoshihiko , Okegawa Takatsugu , Nutahara Kikuo , Hamada Suguru , Nishio Kazuto
Published: April 2023

Distribution of HER4 mutations. The arrows indicate the identified mutations in the HNSCC datasets. The black triangles indicate the previously reported activating mutations, and the red triangle indicates the G1109C mutation. GFR, growth factor receptor; TK, tyrosine kinase.
Supplementary Table 1 from Development of a Novel Antibody–Drug Conjugate for the Potential Treatment of Ovarian, Lung, and Renal Cell Carcinoma Expressing TIM-1
Authors: Thomas Lawrence J. , Vitale Laura , O'Neill Thomas , Dolnick Ree Y. , Wallace Paul K. , Minderman Hans , Gergel Lauren E. , Forsberg Eric M. , Boyer James M. , Storey James R. , Pilsmaker Catherine D. , Hammond Russell A. , Widger Jenifer , Sundarapandiyan Karuna , Crocker Andrea , Marsh Henry C. , Keler Tibor
Published: April 2023

Epitope mapping using overlapping TIM-1 fragments. A series of peptides from the amino acid sequence 209-222 of TIM-1 were synthesized and coated to plates to demonstrate direct binding by CDX-014 mAb.
Supplementary Tables S1 - S6 and Supplementary Figures S1 - S7 from An In Vivo Functional Screen Identifies JNK Signaling As a Modulator of Chemotherapeutic Response in Breast Cancer
Authors: Ashenden Matthew , van Weverwijk Antoinette , Murugaesu Nirupa , Fearns Antony , Campbell James , Gao Qiong , Iravani Marjan , Isacke Clare M.
Published: April 2023

Supplementary Tables (Tables S1 - S6) and Supplementary Figures (Figures S1 - S7)
Supplementary Materials and Methods from Design of a Cytotoxic Neuroblastoma-Targeting Agent Using an Enzyme Acting on Polysialic Acid Fused to a Toxin
Authors: Lehti Timo A. , Pajunen Maria I. , Jokilammi Anne , Korja Miikka , Lilie Hauke , Vettenranta Kim , Finne Jukka
Published: April 2023

Supplementary Materials and Methods describes the procedures for the construction of expression plasmids.
Figure S1 from Design of a Cytotoxic Neuroblastoma-Targeting Agent Using an Enzyme Acting on Polysialic Acid Fused to a Toxin
Authors: Lehti Timo A. , Pajunen Maria I. , Jokilammi Anne , Korja Miikka , Lilie Hauke , Vettenranta Kim , Finne Jukka
Published: April 2023

Figure S1 shows polysialic acid expression levels in neuroblastoma and normal cells.
Figure S4 from Circulating Tumor Cells and Biomarker Modulation with Olaratumab Monotherapy Followed by Olaratumab plus Doxorubicin: Phase Ib Study in Patients with Soft-Tissue Sarcoma
Authors: Martín-Broto Javier , Pousa Antonio López , Brohl Andrew S. , Van Tine Brian A. , Powers Benjamin , Stacchiotti Silvia , Blay Jean-Yves , Hu James S. , Oakley Gerard J. , Wang Hong , Szpurka Anna M. , Levy Donna E. , Mo Gary , Ceccarelli Matteo , Jones Robin L.
Published: April 2023

Representative pre- and post-olaratumab monotherapy biopsies from the same patient, stained with IHC for PDGFRα (A, B) and PDGFRβ (C, D) and displayed at 200x total magnification. Pre-olaratumab images are A and C; post-treatment images are B and D.
Supplementary Table S3 from ER-851, a Novel Selective Inhibitor of AXL, Overcomes Resistance to Antimitotic Drugs
Authors: Tsukamoto Shuntaro , Sugi Naoko Hata , Nishibata Kyoko , Nakazawa Youya , Ito Daisuke , Fukushima Sayo , Nakagawa Takayuki , Ichikawa Kenji , Kato Yu , Kakiuchi Dai , Goto Aya , Itoh-Yagi Machiko , Aota Tomoki , Inoue Satoshi , Yamane Yoshinobu , Murai Norio , Azuma Hiroshi , Nagao Satoshi , Sasai Ken , Akagi Tsuyoshi , Imai Toshio , Matsui Junji , Matsushima Tomohiro
Published: April 2023

Supplementary Table S3
Suppl Figure 4 from Preclinical Characterization of XL092, a Novel Receptor Tyrosine Kinase Inhibitor of MET, VEGFR2, AXL, and MER
Authors: Hsu Jeff , Chong Colin , Serrill Jeffrey , Goon Levina , Balayan Joan , Johnson Eric N. , Lorenzana Grachelle , Wu Sharon , Leong Kevin G. , Yun Theodore J. , Wang Yong , Jiang Faming , Bannen Lynne , Lamb Peter , Xu Wei , Yu Peiwen
Published: April 2023

Suppl Figure 4
Abstract 1901: Analysis of the survivin expression in breast tumor tissue from black and white cancer patients
Authors: Brown Kerrie , Fang Yisheng V. , Sankpal Umesh T.
Published: April 2023

Breast cancer is the most common cancer diagnosed and the leading cause of cancer-related deaths in women. Breast cancer disproportionately affects Black women who have significantly higher mortality rates compared to White women with breast cancer. Although factors like access to healthcare and socioeconomic status contribute significantly to this disparity, there is a need to understand the underlying biological mechanisms that might play an important role. Survivin is a protein that belongs to the inhibitor of apoptosis family. Along with its anti-apoptotic function, it is also involved in the progression of the cell cycle. Survivin is normally expressed during development with no expression detected in most adult differentiated tissues. However, high expression of this protein is found in many cancers which correlates with poor prognosis. The goal of this study was to determine the expression of survivin in tumor tissue from Black and White breast cancer patients. Our first approach was to analyze the TCGA database to confirm our hypothesis that there is differential expression of survivin between the two racial groups and within different subtypes of breast cancer. Secondly, we used immunohistochemical analysis to investigate the expression of survivin in patient tumor tissue samples obtained from Black and White breast cancer patients. Results from the analysis of the TCGA database indicate significantly higher expression of survivin in tumors from Black patients compared to White. High survivin expression is also correlated with higher mortality rates. Immunohistochemical analysis of breast tumors also demonstrated higher expression of survivin in tumor tissue from Black patients. We also observed higher nuclear expression in Black patients compared to White patients. Differential expression was also observed between different subgroups of breast cancer. High expression was seen in the most aggressive triple negative (TNBC) form of breast cancer, which is known to disproportionately occur at higher rates in Black women. Our results along with data available in literature, suggest that survivin plays an important role in breast cancer progression (in correlation to race) and could be a useful diagnostic and/or prognostic marker or a therapeutic target.

Citation Format: Kerrie Brown, Yisheng V. Fang, Umesh T. Sankpal. Analysis of the survivin expression in breast tumor tissue from black and white cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1901.
Abstract 2578: Phosphorylation of AHR by PLK1 promotes metastasis of LUAD via upregulation of DIO2
Authors: Li Chaohao , He Daheng , Mao Fengyi , Wang Xinyi , Hao Yanning , Kong Yifan , Brainson Christine F. , Liu Jinghui , Zhang Yanquan , Wang Ruixin , Zhang Qiongsi , Li Zhiguo , Rao Xiongjian , Wu Sai , Wang Chi , Wei Qiou , Wang Jianlin , Liu Xiaoqi
Published: April 2023

Non-small cell lung cancer (NSCLC) is one of the most aggressive cancer types, and metastasis of cancer cells is the major reason leading to death of patients. Aryl hydrocarbon receptor (AHR) is an important transcription factor involved in the initiation and progression of lung cancer. Polo-like kinase 1 (PLK1), a serine/threonine kinase, is another oncogene that promotes the malignancy of multiple cancer types. Preliminary examination indicates that AHR is a potential substrate of PLK1. Nonetheless, the phosphorylation and interaction of these two factors, as well as the subsequent biological significance in lung cancer remain to be determined. Here we prove that PLK1 phosphorylates AHR at serine 489 (S489) in lung adenocarcinoma (LUAD), a subtype of NSCLC. Overexpressing of exogenous AHR with mutation of phosphomemetic amino acid (S489D), enhances migration and invasion in transwell assays compared to cells harboring AHR that is unable to be phosphorylated (S489A), indicating that phosphorylation of AHR by PLK1 induces the metastatic potential of lung cancer. This is further confirmed in mouse studies using intravenous or subcutaneous inoculation of cancer cells. RNA-seq analyses of cells with S489D or S489A show that type 2 deiodinase (DIO2) is underrepresented in S489A but overexpressed in S489D. DIO2 converts thyroxine (T4) to triiodothyronine (T3), which is the more active form of thyroid hormone and activates downstream effect. Treatment with T3 or T4 promotes the metastasis of LUAD, whereas depletion of DIO2 or treatment with deiodinase inhibitor iopanoic acid disrupts this property. Furthermore, this phenomenon is verified by in vivo mouse experiment using intravenous injection of DIO2-depleted cancer cells. Taken together, these results identify phosphorylation of AHR by PLK1 as a mechanism that leads to progression of LUAD.

Citation Format: Chaohao Li, Daheng He, Fengyi Mao, Xinyi Wang, Yanning Hao, Yifan Kong, Christine F. Brainson, Jinghui Liu, Yanquan Zhang, Ruixin Wang, Qiongsi Zhang, Zhiguo Li, Xiongjian Rao, Sai Wu, Chi Wang, Qiou Wei, Jianlin Wang, Xiaoqi Liu. Phosphorylation of AHR by PLK1 promotes metastasis of LUAD via upregulation of DIO2 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2578.
Abstract 287: Novel p62 interaction suggests novel role in glutamate metabolism
Authors: Dahl Haley Christine , Konakalla Anisha R. , Kanumuri Preethi , Mansoor Basir , Kelley Anastasia , Shipurkar Riya , Delk Nikki A.
Published: April 2023

Prostate cancer (PCa) is the second most common cause of cancer-related death among American men. Androgen Receptor (AR) transcriptional activity is required for PCa tumor growth. Thus, androgen deprivation therapy (ADT) is a first line PCa therapy. However, 10-20% of patients will inevitably develop lethal castration-resistant PCa (CRPC). Thus, we must uncover the mechanisms that drive CPRC. Chronic inflammation is a known hallmark of cancer initiation and progression. Using the LNCaP human PCa cell line, we have shown that chronic exposure to the inflammatory cytokine, interleukin-1 (IL-1) can transform castration-sensitive PCa cells into castration-resistant PCa cells. IL-1 is elevated in PCa patient tissue and serum and is associated with disease progression and metastasis. IL-1 is clinically relevant, but the role of IL-1 in CRPC development is not fully elucidated. To mimic ADT, we serum-starved LNCaP and the chronic IL-1 sublines, LNas1 and LNbs1, and performed mass spectrometry to identify chronic IL-1-induced changes in protein-protein interactions. Specifically, we looked at p62 protein interactors. p62 is a multi-domain, multifunctional pro-survival protein that mediates autophagic flux in response to cellular stress and serum starvation, promotes NF-κB inflammatory signaling and induces NRF2 antioxidant signaling. We discovered that p62 forms a constitutive interaction with KEAP1 under serum starved and normal growth conditions in the chronic IL-1 sublines. KEAP1 inhibits NRF2 nuclear translocation. In response to antioxidant stress, p62 binds and sequesters KEAP1 from NRF2 to activate NFR2 signaling. Surprisingly, we do not observe constitutive canonical NRF2 signaling in the chronic IL-1 sublines. However, in response to serum starvation, we do see both high basal levels and upregulation, specifically, of the NRF2-induced gene, GCLC. GCLC is a glutamate-cysteine ligase that synthesizes the antioxidant, glutathione. Therefore, using mass spectrometry and immunoprecipitation western blotting, we looked at p62 interactions under serum starvation conditions. While we did not find p62 interaction with GCLC in the chronic IL-1 sublines, we do find that p62 interacts with GLUL. GLUL is a glutamate-ammonia ligase that plays a role in cellular detoxification. These data suggests that chronic IL-1 exposure induces novel p62 interactions that have a role in glutamate metabolism.

Citation Format: Haley Christine Dahl, Anisha R. Konakalla, Preethi Kanumuri, Basir Mansoor, Anastasia Kelley, Riya Shipurkar, Nikki A. Delk. Novel p62 interaction suggests novel role in glutamate metabolism [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 287.
Abstract 2945: Depleting hCCR8 mAb Therapy #2: Selection of candidates for the development of innovative depleting anti CCR8 therapeutic antibodies to control the immunosuppressive tumor microenvironment
Authors: Maurin Alice Gentil Dit , Richert Iseulys , Fernandez Maria Dolores Garcia , Janvier Aurélie , Rose Solene , Jeannelle Megane , Chraa Dounia , Dietsch Camille , Urquia Pauline , Ruet Quentin , Schappler Malaury , Baron Luc , Ayachi Safia , Blanchard Orphée , Franchet Christel , Fontayne Alexandre , Vermot-Desroches Claudine , Schann Stephan , Lenne Nathalie
Published: April 2023

Tumor-infiltrating Tregs (TITR) constitute a sub-family of immuno-suppressive cells abundantly found in multiple solid cancers. They play a critical role in tumor progression and their specific depletion has been recently propose as a novel therapeutic strategy to fight cancers. Based on the transcriptional analysis of tumor infiltrating immune cells, the G-protein coupled receptor CCR8 was found to be expressed on murine and human TITR, but not on proinflammatory effector T cells. The CCR8 receptor therefore represents a unique TITR target for the development of novel depletive antibody-based therapeutics. At Domain Therapeutics, a diverse library of several dozens of mouse mAbs directed against the hCCR8 was discovered and characterized, providing a unique source for the development of a best-in-class and well differentiated anti-hCCR8 depleting antibody for the treatment of cancers.Special attention was paid on two important characteristics for the selection of our depleting mAb candidates: (1) their ability to detect all CCR8 positive immunosuppressive cells and (2) the combination of different killing modes of action (ADCC, ADCP, CDC) to trigger an optimal depletion.Our selected mAb candidates were nominated based on their capacity to bind to immunosuppressive cells present in different tumor tissues and optimal depleting activity. The impact of our mAbs on tumor growth and on the immune tumor microenvironment (TME) were also studied in mouse hCCR8-KI mice. Finally, following a comprehensive in vitro benchmarking mAb analysis, we identified key differentiating points versus competitor’s mAbs. Nominated candidates wil be ready to enter cell line generation efforts in early 2023.

Citation Format: Alice Gentil Dit Maurin, Iseulys Richert, Maria Dolores Garcia Fernandez, Aurélie Janvier, Solene Rose, Megane Jeannelle, Dounia Chraa, Camille Dietsch, Pauline Urquia, Quentin Ruet, Malaury Schappler, Luc Baron, Safia Ayachi, Orphée Blanchard, Christel Franchet, Alexandre Fontayne, Claudine Vermot-Desroches, Stephan Schann, Nathalie Lenne. Depleting hCCR8 mAb Therapy #2: Selection of candidates for the development of innovative depleting anti CCR8 therapeutic antibodies to control the immunosuppressive tumor microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2945.
Abstract 3442: FXR mediates macrophage intrinsic responses to suppress colon cancer progression
Authors: Fu Ting , Dong Xingchen
Published: April 2023

Bile acids (BAs) affect the intestinal environment by ensuring barrier integrity, regulating epithelium turnover, maintaining microbiota balance, and modulating the immune system. As a master regulator of BAs homeostasis and innate immunity, Farnesoid X Receptor (FXR), is severely compromised in inflammatory bowel disease (IBD) and colitis-associated colorectal carcinoma (CAC) patients. At the front line, gut macrophages react to the microbiota and metabolites that breach the epithelium. We aim to study the role of the BAs-FXR axis in macrophages. To dissect how inflammation drives tumorigenesis, we employed a classic mouse model of CAC, profiled BAs and cytokines, monitored tumorigenesis, and examined the function of macrophages and Th17 cells. Moreover, we investigated the potential of FexD, an FXR agonist, in ameliorating intestinal inflammation, thus inhibiting tumorigenesis. Further, we identified how macrophage intrinsic FXR senses the BAs abnormality and reshapes the gut macrophages in vivo and ex vivo. We also investigated the potential role of pro-inflammatory cytokines in stimulating intestinal stem cells (ISCs)’ stemness using mouse organoids. This study demonstrates that inflammation-induced epithelial abnormalities compromised FXR signaling and altered BAs profile in CAC. Gut macrophage intrinsic FXR senses the aberrant BAs, leading to the secretion of pro-inflammatory cytokines, which promotes ISCs’ proliferation. Notably, FexD reduced intestinal inflammation and tumorigenesis by suppressing pro-inflammatory responses in gut macrophages and Th17 cells. Mechanistically, FXR regulates gut macrophages' recruitment, polarization, maturation, and crosstalk with Th17 cells. In summary, we uncovered a novel role of FXR modulation of gut macrophages, highlighting the potential of FXR as a therapeutic target over biologicals for treating colon cancer.

Citation Format: Ting Fu, Xingchen Dong. FXR mediates macrophage intrinsic responses to suppress colon cancer progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3442.
Abstract 3497: The polyclonal path to malignant transformation in familial adenomatous polyposis
Authors: Schenck Ryan O. , Khan Aziz , Horning Aaron , Esplin Edward D. , Monte Emma , Wu Si , Hanson Casey , Bararpour Nasim , Neves Stephanie , Jiang Lihua , Contrepois Kévin , Lee Hayan , Guha Tuhin K. , Hu Zheng , Laquindanum Rozelle , Mills Meredith A. , Chaib Hassan , Chiu Roxanne , Jian Ruiqi , Chan Jeannie , Ellenberger Mathew , Becker Winston R. , Bahmani Bahareh , Michael Basil , Shen Jeanne , Lancaster Samuel , Ladabaum Uri , Kundaje Anshul , Longacre Teri A. , Greenleaf William J. , Ford James M. , Snyder Michael P. , Curtis Christina
Published: April 2023

Familial adenomatous polyposis (FAP) patients develop hundreds of premalignant polyps that progress to colorectal cancer due to a germline mutation in the APC tumor suppressor. Polyps from FAP patients uniquely facilitate interrogation of the continuum of malignant transformation from histologically normal mucosa to benign and dysplastic polyps and eventual adenocarcinomas. As part of the Human Tumor Atlas Network (HTAN), we performed multi-omic profiling, including whole genome sequencing, on 135 samples from six FAP patients across the pre-malignant continuum spanning all physical regions of the large intestine, and serving as the most comprehensive FAP dataset available. Through a comparative analysis with a published FAP cohort (58 multi-region samples across 5 patients) and sporadic colorectal cancer cohort (n=57), each including benign and malignant samples, we evaluate the timing of driver acquisitions at each stage of malignant progression. Despite being separated by vast regions of histologically normal mucosa, independently evolving polyps show extensive mutation sharing, suggesting FAP polyps are polyclonal in origin. Finally, we leverage a simplified mechanistic model of embryonic colonic development demonstrating that the path to malignant transformation in FAP is consistent with polyclonal origins attributable to early mixing, perhaps as early as in-utero. Taken together, the HTAN FAP Atlas provides a novel window into the earliest stages of cancer formation and may illuminate barriers to malignant transformation and opportunities for earlier intervention.

Citation Format: Ryan O. Schenck, Aziz Khan, Aaron Horning, Edward D. Esplin, Emma Monte, Si Wu, Casey Hanson, Nasim Bararpour, Stephanie Neves, Lihua Jiang, Kévin Contrepois, Hayan Lee, Tuhin K. Guha, Zheng Hu, Rozelle Laquindanum, Meredith A. Mills, Hassan Chaib, Roxanne Chiu, Ruiqi Jian, Jeannie Chan, Mathew Ellenberger, Winston R. Becker, Bahareh Bahmani, Basil Michael, Jeanne Shen, Samuel Lancaster, Uri Ladabaum, Anshul Kundaje, Teri A. Longacre, William J. Greenleaf, James M. Ford, Michael P. Snyder, Christina Curtis. The polyclonal path to malignant transformation in familial adenomatous polyposis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3497.
Abstract 3737: A novel mechanism of NF-κB activation by genotoxic chemotherapy
Authors: Venkatesh Jaganathan , Muthu Magesh , Rishi Arun K.
Published: April 2023

CARP-1, a perinuclear phospho-protein, is a biphasic regulator of cell survival and apoptosis signaling. Genotoxic drugs induce apoptosis in part by elevating CARP-1 levels while also promote cell survival in part by activating NF-κB that involves CARP-1 interaction with NF-κB kinase subunit γ (aka, NEMO). We previously noted interaction of CARP-1 with RIPK1, and since RIPK1 is known to regulate NF-κB signaling, we investigated whether RIPK1-CARP-1 interaction is involved in genotoxic chemotherapy-dependent apoptosis or NF-κB-activated survival signaling. Mutagenesis and co-IP-WB analyses revealed that a 40-amino acid epitope within the amino-terminal catalytic domain of RIPK1 interacted with CARP-1 (611-640) peptide. Overexpression of this CARP-1/CCAR1-binding (CB) epitope peptide or RIPK1 mutant that has in-frame deletion of CB-epitope resulted in moderate but significant reduction in viabilities of cells treated with Adriamycin or Cisplatin relative to the viability loss noted in cells expressing RIPK1 (WT). However, no significant differences in loss of cell viabilities were noted in cells expressing various substitution mutants of RIPK1 (aspartate (D) 138 to asparagine (N), serine (S) 109 and threonine (T) 110 to alanines (A), lysine (K) 87 to arginine (R), K97 to R, K105 to R, or K115 to R) relative to their RIPK1 (WT) expressing counterparts when treated with genotoxic drugs. Interestingly, CB-epitope mutant, but not RIPK1 (WT), failed to translocate to cytoplasm in Adriamycin-treated cells. Genotoxic drugs activated p65/RelA in cells expressing RIPK1 (WT), while expression of RIPK1 CB-epitope mutant resulted in abrogation of p65/RelA activation by Adriamycin or Cisplatin but not by TNFα. Collectively our data demonstrate that CARP-1 interaction with RIPK1 promotes nuclear export of RIPK1 and consequent NF-κB activation in cells exposed to genotoxic drugs.

Citation Format: Jaganathan Venkatesh, Magesh Muthu, Arun K. Rishi. A novel mechanism of NF-κB activation by genotoxic chemotherapy. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3737.
Abstract 3739: MAFG promotes melanomagenesis through the transcriptional regulation of hypoxic and immune responses
Authors: Vera Olga , Martinez Michael , Karreth Florian A.
Published: April 2023

The small MAF family of transcription factors, consisting of MAFG, MAFF, and MAFK, dimerize with CNC/BACH proteins to regulate diverse transcriptional programs. Previously, we have shown that miRNA-29 suppresses melanoma development, at least in part, by repressing MAFG expression. Moreover, analysis of the TCGA revealed frequent copy number gains and/or overexpression of MAFG in melanoma. However, whether MAFG has oncogenic effects in and is critical for melanoma is unknown. We found that silencing of MAFG, but not MAFF or MAFK, abrogates growth of melanoma cells in vitro, while melanocyte cell lines are less reliant on MAFG. Conversely, MAFG overexpression promoted proliferation and focus formation of human melanocyte and melanoma cell lines. Using a high-throughput melanoma mouse modeling (ESC-GEMM) platform, we found that in vivo overexpression of MAFG in BRAFV600E; PTEN+/- mice potently accelerated melanoma development. RNA-sequencing revealed that MAFG regulates transcriptional programs associated with hypoxic and immune responses. Accordingly, MAFG overexpressing cells exhibited better ability to survive under hypoxic conditions, probably through the regulation of HIF1A. Moreover, MAFG overexpressing cells showed increased expression of cytokines of the of the C-C chemokine family, CCL3 and CCL5. MAFG overexpressing tumors from ESC-GEMM mice and MAFG-High melanomas from TCGA had features of immune-cold milieus, characterized by reduced infiltration of CD45+ immune cells and CD8+ T cells. Altogether, we demonstrate that MAFG promotes melanomagenesis, in part, by regulating the response to hypoxic conditions and chemokine expression, which may rewire the tumor immune microenvironment.

Citation Format: Olga Vera, Michael Martinez, Florian A. Karreth. MAFG promotes melanomagenesis through the transcriptional regulation of hypoxic and immune responses. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3739.
Abstract 387: Enolase-1 as a candidate theranostics target for neuroendocrine prostate cancer
Authors: Torres Krystal Rubi Santiago , Duran Alfonso
Published: April 2023

Prostate cancer (PCa) is the second most common cancer in American men, with men of African ancestry (AA) having twice the mortality of men of European ancestry (EA). Although taxane-based chemotherapy is the last line of defense in men with advanced PCa, it ultimately fails due to chemoresistance. The protein-specific membrane antigen (PSMA) has been an effective target for the imaging and therapy of advanced PCa. Specifically, PSMA radioligand therapy (PSMA-RLT) is a theranostics (therapy + diagnostics) option for men with advanced PCa. However, about 30% of men with advanced PCa have limited response to PSMA-RLT due to the presence of neuroendocrine-differentiated PCa (NEPC), which lacks PSMA expression. A promising alternative to PSMA targeting is the glycolytic enzyme enolase (ENO), which localizes to the cell surface in advanced tumors. There are three enolases with similar functions, with ENO-1 and ENO-2 implicated in PCa. Here, we demonstrate, using immunoblotting, confocal microscopy, and cell fractionation studies with specific monoclonal antibodies, that chemosensitive NEPC cells express both ENO-1 and ENO-2; however, docetaxel (DTX)-resistant PC3-DR and DU145-DR cells only express ENO-1 with clear loss of ENO-2. Loss of ENO-2 in chemoresistant PCa cells generates a metabolic vulnerability due to loss of ENO redundancy. We also initiated the testing of small molecule inhibitors (SMIs) targeting ENO-1 in chemoresistant and chemosensitive NEPC cells, showing that these inhibitors have cytotoxic activity against these cells, with IC50 values in the low micromolar range. These results support the hypothesis that ENO-1 is expressed in the surface of NEPC cell lines and can be targeted with SMIs that could be modified as potential theranostics agents. Our long-term goal is to identify an alternative treatment for patients with NEPC by establishing ENO-1 as a novel cell surface theranostics target.

Citation Format: Krystal Rubi Santiago Torres, Alfonso Duran. Enolase-1 as a candidate theranostics target for neuroendocrine prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 387.
Abstract 3959: Novel spliced isoform of the proteasome subunit ADRM1/Rpn13 promotes hepatocellular carcinoma (HCC) development through selective degradation of tumor suppressor p53
Authors: Sun Yanmei , Xu Mingjing , Wan Ho Lee , Wong Alissa M. , Ng Kelvin K. , Wong Nathalie
Published: April 2023

The Ubiquitin-Proteasome System (UPS) is the major pathway in eukaryotic cells that regulates intracellular protein degradation and turnover. Cumulative evidence has suggested that defects in the UPS machinery are involved in the pathogenesis of human diseases, including cancer. The proteasome-associated polyubiquitin receptor ADRM1 (also known as Rpn13) orchestrates the process of substrate recruitment with deubiquitination in the proteasome through its N-terminal ubiquitin-binding domain and C-terminal activation of deubiquitinating enzyme. Using long-read SMRT-seq to profile the RNA splicing landscape in hepatocellular carcinoma (HCC), we previously reported on an unannotated alternative spliced variant of ADRM1 in human hepatocellular carcinoma (HCC) (Hepatology 2019). This novel isoform exhibits an exon 9 skipping (ADRM1-ΔEx9) that resulted in an altered C-terminus of the protein. In this study, we undertook further investigations to characterize the detailed functional effects of ADRM1-ΔEx9 in HCC biology and the underlying mechanism. Initial study by junction-specific Taqman PCR assay showed that ADRM1-ΔEx9 is more frequently upregulated in HCC tumors than the canonical full-length counterpart (ADRM1-FL). There is an interchange between the two isoforms as tumor develops which underscores an isoform-switch in favor of HCC development. Indeed, our result showed that ADRM1-ΔEx9, but not ADRM1-FL, correlated significantly with inferior patient survival. Functionally, ADRM1-ΔEx9 knockdown profoundly suppressed proliferation by inducing spontaneous apoptosis in HCC cell lines and patient-derived HCC organoids. The corresponding effects were less apparent from ADRM1-FL knockdown. Consistently, overexpression of ADRM1-ΔEx9 in normal human liver organoids promoted pre-malignant features alongside increased in-vitro propagation time. Ubiquitin Proteome profiling revealed that ADRM1-ΔEx9 led to a reduced expression of the pivotal tumor suppressor p53. A parallel transcriptome also substantiated that ADRM1-ΔEx9 modulated genes involved in the p53 and apoptosis pathways. Collectively, our study demonstrated that the non-canonical spliced isoform ADRM1-ΔEx9 confers growth and pro-survival advantages in HCC through mediating selective degradation of tumor suppressor p53 protein.

Citation Format: Yanmei Sun, Mingjing Xu, Ho Lee Wan, Alissa M. Wong, Kelvin K. Ng, Nathalie Wong. Novel spliced isoform of the proteasome subunit ADRM1/Rpn13 promotes hepatocellular carcinoma (HCC) development through selective degradation of tumor suppressor p53. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3959.
Abstract 4006: TRX-221, a novel 4th-generation EGFR inhibitor for overcoming C797S mutation-mediated acquired resistance in NSCLC
Authors: Lim Sumin , Choi Seulgi , Lee Ju-Kyung , Choi Eunhyun , Lee Sungwon , Chun Kwangwoo , Baek Myoungki , Park Jiyeon , Kim Jihyun , Kang Areum , Kim Younghoon , Kim Namkyoung , Cho Byoungchul , Sim Taebo , Lee Koo
Published: April 2023

Targeting mutated EGFR is a clinically validated approach for tackling non-small cell lung cancer (NSCLC). Approximately 20% of patients with advanced NSCLC harbor an EGFR mutation. Osimertinib is a 3rd-generation oral EGFR-TKI targeting the EGFR sensitizing mutations Del19 and L858R, as well as an acquired mutation, T790M. Despite osimertinib's efficacy in NSCLC, resistance develops and the mechanisms of resistance to osimertinib are complex. Development of another mutation, C797S is the most common mechanism of acquired resistance following osimertinib treatment. This mutation occurs with about 12% frequency after a 1L therapy of osimertinib, and with approximately 17% frequency after a 2L therapy. Here, we report TRX-221 as a novel 4th-generation EGFR-TKI that can overcome C797S-mediated acquired resistance in NSCLC models in vitro and in vivo. TRX-221 exhibited potent inhibitory activity against multiple EGFR mutant kinases, including Del19/T790M/C797S, L858R/T790M/C797S, Del19/C797S, L858R/C797S, Del19/T790M, L858R/T790M, Del19, and L858R. Likewise, TRX-221 potently inhibited the proliferation of Ba/F3 cells stably overexpressing these eight EGFR mutants. TRX-221 however showed sparing activity against EGFR wild-type kinase and cell lines. TRX-221 also demonstrated marked anti-proliferative activity against osimertinib-resistant patient-derived cancer cell lines (PDCs). With favorable DMPK profiles, once-daily oral dosing of TRX-221 induced strong anti-tumor effects in a dose-dependent manner in a variety of osimertinib-resistant CDX and PDX models. As a reversible, selective, and broad-spectrum 4th-generation EGFR-TKI, TRX-221 is currently undergoing IND-enabling studies and shows strong potential as a therapeutic solution to treat advanced NSCLC patients harboring EGFR mutations.

Citation Format: Sumin Lim, Seulgi Choi, Ju-Kyung Lee, Eunhyun Choi, Sungwon Lee, Kwangwoo Chun, Myoungki Baek, Jiyeon Park, Jihyun Kim, Areum Kang, Younghoon Kim, Namkyoung Kim, Byoungchul Cho, Taebo Sim, Koo Lee. TRX-221, a novel 4th-generation EGFR inhibitor for overcoming C797S mutation-mediated acquired resistance in NSCLC. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4006.
Abstract 4068: Ganglioside SSEA4 in Ewing sarcoma: A marker of tumor cells with highly aggressive features and a potential immune target
Authors: Jamitzky Silke , Altvater Bianca , Krekeler Carolin , Hoen Laura , Brandes Caroline , Richter Lisa , Ebbinghaus Julia , Ochs Laurin , Farwick Nicole , Urban Katja , Görlich Dennis , Johnston Ian , Pfeifer Rita , Rossig Claudia , Hartmann Wolfgang , Kailayangiri Sareetha
Published: April 2023

Cancers can aberrantly express carbohydrate antigens restricted to immature cells during prenatal human development, and these may be exploited for selective immune targeting. We studied expression and functional associations of the globo-series ganglioside stage-specific embryonic antigen 4 (SSEA4), a cell surface marker of human embryonic stem cells, in Ewing sarcoma (EwS). Flow cytometry revealed SSEA4 expression on the cell surface of each of 13 EwS cell lines, with moderate to high expression densities in 6 of 13 (46%) cell lines. Of 31 primary EwS tumor biopsies, 21 (68%) were SSEA4 positive by immunofluorescence staining. No associations of SSEA4 positivity with molecular or clinical disease parameters were found. Comparisons of paired tumor cell subpopulations with high and low/negative SSEA4 expression selected by cell sorting from 8 EwS cell lines revealed increased cell proliferation and colony formation of SSEA4high tumor cells, along with higher doxocrubicin chemoresistance and a higher propensity to migrate in transwell assays. SSEA4 negative cells selected from bulk populations of EwS cells regained SSEA4 expression during continued in vitro culture as well as after in vivo xenografting into immunodeficient mice. SSEA4high expression in EwS cells was not dependent on expression levels of the EWSR1-FLI1 fusion transcript or its direct targets, nor associated with markers of epithelial/mesenchymal plasticity. To retarget human T cells to SSEA4, we generated a second-generation, 41BBζ CAR using the antigen-binding domains of a murine anti-SSEA4 monoclonal antibody. SSEA4-specific CAR T cells specifically interacted with SSEA4 positive EwS cells in a strictly antigen-dependent manner, resulting in effective tumor cell lysis in vitro. We conclude that targeting of SSEA4 with CAR T cells or alternative immune therapeutics could be an attractive strategy to eliminate tumor cell subsets with high propensity to drive disease progression in EwS and therefore deserves further evaluation towards clinical translation.

Citation Format: Silke Jamitzky, Bianca Altvater, Carolin Krekeler, Laura Hoen, Caroline Brandes, Lisa Richter, Julia Ebbinghaus, Laurin Ochs, Nicole Farwick, Katja Urban, Dennis Görlich, Ian Johnston, Rita Pfeifer, Claudia Rossig, Wolfgang Hartmann, Sareetha Kailayangiri. Ganglioside SSEA4 in Ewing sarcoma: A marker of tumor cells with highly aggressive features and a potential immune target. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4068.
Abstract 4097: Functional and translatable efficacy of CAR-T cells targeting B cell lymphomas
Authors: Kelsey Lauren , Latta Karin , Kolb Joey , John Bincy , Aylott Ilona , Rocca Daniel , Bacita Alexandru , Kirkham Christopher , Walters Patrick , Humphery Charlotte , Ballesteros Lorena Sueiro , Self-Fordham Jezrom , Brackenbury Louise , Hall Chassidy , David Harris , Schueler Julia , Nunan Robert
Published: April 2023

Chimeric Antigen Receptor (CAR)-T cell therapy has achieved great success in treating a variety of liquid tumors, in particular CD19+ lymphomas. CAR technology is advancing rapidly, with notable improvements in efficacy, safety and complexity as human ingenuity seeks solid tumor applicability. Here we describe how to efficiently generate, expand and validate in-vitro and in-vivo efficacy of a clinically tested CD19-specific CAR. HLA-A*02:01+ CD3+ T-cells were transduced to express a CD19-CAR to >80% efficiency (in 5 donors) and reproducibly expanded to >68 fold over a 19 day period. In-vitro studies were used to assess tumor killing ability, utilizing a HLA-A*02:01+ patient derived xenograft (PDX) CD19+ cell line, alongside human cancer cell lines RAJI (CD19+) and K562 (CD19-). Analysis revealed the potent ability of CD19-CAR-T cells to specifically target CD19+ cells in a dose dependent manner. Supernatant analysis showed highly elevated IFNγ concentrations, identifying enhanced activation of CAR-T cells in the presence of CD19+ target cells only. In-vivo survival studies were completed to identify translatability & efficacy in both systems. Mice were inoculated with CD19+ cells to produce multiple tumor burdened models (RAJI, JEKO-1 & PDX4009). This demonstrated high efficacy of CAR-T at targeting both tumor cell line RAJI or PDX in vivo, with almost complete tumor regression seen in both models. JEKO-1 tumors saw progression slowed in comparison to non-transduced or no treatment controls however due to the nature of this cell line (highly tumorigenic) the level of tumor regression was not matched to the other models. Cell bio-distribution was assessed with CAR-T cells tracking successfully to the tumors as well as the highly vascularized tissues, which was expected and consistent with previous data. These assays can be translated to any CAR construct aimed at targeting either solid or hematological tumors and demonstrates an integrated platform for translating CAR research that can be utilized end to end.

Citation Format: Lauren Kelsey, Karin Latta, Joey Kolb, Bincy John, Ilona Aylott, Daniel Rocca, Alexandru Bacita, Christopher Kirkham, Patrick Walters, Charlotte Humphery, Lorena Sueiro Ballesteros, Jezrom Self-Fordham, Louise Brackenbury, Chassidy Hall, Harris David, Julia Schueler, Robert Nunan. Functional and translatable efficacy of CAR-T cells targeting B cell lymphomas. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4097.