bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1780: Utilizing bisbiguanide compounds to disrupt metabolic heterogeneity in non-small cell lung cancer
Authors: Matsuk Veronika Y. , Knippler Christina M. , Arnst Jamie L. , Robinson Isaac E. , Khatib Tala O. , Shanmugam Mala , Ganesh Thota , Mouw Janna K. , Marcus Adam I.
Published: March 2024

Metastasis accounts for 90% of cancer-related deaths; however, the molecularmechanisms by which cancer cells invade and metastasize remain poorly understood.Around 80% of lung cancer patients present with metastatic disease and have only a 5%5-year relative survival. Metastases are often seeded by heterogeneous cells that invadecollectively in cellular packs. However, most studies and treatments focus on bulk cellularpopulations leaving subpopulations that drive metastasis masked. We previouslypublished on metabolic heterogeneity within collectively invading lung cancer cells andidentified the bisbiguanide compound alexidine as a disrupter of lung cancermitochondrial metabolism and invasion. To expand on these findings, we developed newbiguanides and identified promising candidates, AX-4 and AX-7. These analogs inducemitochondrial structural defects and cause metabolic rewiring. Treated lung cancer cellsexhibited reduced oxygen consumption and increased lactate production. Importantly,AX-4 and AX-7 reduced collective cell invasion, highlighting the potential of these newbiguanides to limit cancer metastasis. Taken together, we have developed new chemicaltools to disrupt mitochondrial metabolism. These tools will be useful to further parse theroles of metabolically heterogenous cancer cells on tumor growth and metastasis.

Citation Format: Veronika Y. Matsuk, Christina M. Knippler, Jamie L. Arnst, Isaac E. Robinson, Tala O. Khatib, Mala Shanmugam, Thota Ganesh, Janna K. Mouw, Adam I. Marcus. Utilizing bisbiguanide compounds to disrupt metabolic heterogeneity in non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1780.
Abstract 179: Identification of a novel crosstalk between MDSC and NKregs in immunotherapy resistant triple negative breast cancer
Authors: Das Megha , Henry Samantha , Singh Shailesh , Santos Camila Dos , Chakrabarti Rumela
Published: March 2024

Triple-negative breast cancer (TNBC) is one of the most aggressive forms of breast cancer and lacks standard treatment options. Although immunotherapy shows some response in these patients along with chemotherapy, heterogeneity of tumor cells and immune cells results in significant low response to immunotherapy treatment. Based on single cell sequencing which enables identity of heterogenous population at single cell level and functional studies involving preclinical mouse models, our laboratory recently identified distinct CD56brightSOCS3highCD11b−CD27− immature NK cells subpopulations, with diminished cytotoxic granzyme signatures that promote TNBC tumor progression. However, the precise characterization and functionality of these rogue tumor promoting NK cells remains unclear in clinical samples. Here, we aimed to illustrate mechanistic insight of these novel immature NK subpopulation in context of immunotherapy resistant TNBC patient samples. The scRNA seq analysis of TNBC patient samples identified four specific subclusters of NK cells in TNBC. We further found that NK2 and NK3 subclusters are CD56dim Granzymehigh cytotoxic NK cells and the other two subclusters (NK0 and NK1) are CD56bright immature NKs. Interestingly, the NK0 and NK1 subcluster specifically exhibit distinctive characteristics and functions. Notably, CD56bright immature NK1 cells, identified as tissue-resident NK cells, were engaged in direct communication with myeloid-derived suppressor cells (MDSCs). The CD56bright NK0 possesses moderate to low SOCS3 expression and moderate to high granzyme expression with elevated early NK activation markers and inhibitory receptor expressions, suggesting its regulatory property (NKregs). scRNA seq and flow cytometry analysis showed higher NKregs and immunosuppressive MDSC population, specifically with high monocytic-MDSCs (mMDSCs) in non-responder TNBC (to adjuvant combinational treatment of chemo and immunotherapy) patient samples rather than responder patients’ tissue. Based on in silico analysis, we further found a potential crosstalk between these mMDSC & NKreg within the tumors of non-responsiveness of TNBC patients. Overall, our studies show a connection of MDSCs and NKregs in TNBC patients, who do not respond to immunotherapy, highlighting a possible mechanism for immunotherapy resistance, which could be targeted in near future for better treatment options in TNBC.

Citation Format: Megha Das, Samantha Henry, Shailesh Singh, Camila Dos Santos, Rumela Chakrabarti. Identification of a novel crosstalk between MDSC and NKregs in immunotherapy resistant triple negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 179.
Abstract 1843: Integrating machine learning with microfluidic technologies for proteomic profiling of extracellular vesicles in triple-negative breast cancer
Authors: Kim Min Woo , Kim Jee Ye , Kim Young , Lee Suji , Moon Sol , Hyon Ju-yong , Hyun Kyung-A , Yang Yeji , Ha Seongmin , Park Sunyoung , Gawk Hogyeong , Lee Haeji , Han Eun Hee , Kim Jin Young , Jung Hyo-Il , Chung Young-Ho , Kim Seung Il
Published: March 2024

Accurate diagnosis of breast cancer using circulating biomarkers present in plasma remains an important challenge. In particular, protein changes in tumor-derived extracellular vesicles (tdEVs) have emerged as potential biomarkers for breast cancer diagnosis because they accurately reflect dynamic changes in tumors. In this study, we compared the proteomes of extracellular vesicle (EV) isolated from the plasma of 100 breast cancer patients and 30 healthy individuals who visited Severance Hospital between Mar 2010 and Dec 2015. Microfluidic chip-based protocol that facilitates the removal of contaminants such as albumin and immunoglobulins was used for the extraction of enriched tdEVs with small amounts of plasma. Comparative analysis of the proteomes identified 26 significant biomarkers that could indicate differences between breast cancer patients and healthy individuals. Using the LsBoost-CNN-SVM hybrid machine learning algorithm, we especially identified key EV protein biomarkers for detecting triple-negative breast cancer (TNBC), a breast cancer subtype with a poor prognosis and a high recurrence rate, as well as lacking therapeutic and diagnostic targets. Remarkably, a signature consisting of three EV proteins, specifically extracellular matrix protein 1 (ECM1), mannose-binding lectin 2 (MBL2), and biotinidase (BTD), effectively distinguished TNBC patients from healthy individuals. In our proteomic analysis set (n=73), this signature exhibited impressive performance, achieving a sensitivity of 93.3% and specificity of 93% in the accurate discrimination of TNBC from the control group. The validation set (n=40) confirmed these findings, with 100% sensitivity and 80% specificity. This signature not only served as a diagnostic tool but also provided valuable insights into the risk of recurrence and patient prognosis. We found a novel diagnostic approach that holds the potential to revolutionize breast cancer diagnostics by enhancing the reliability of tumor-related information obtained from blood samples.

Citation Format: Min Woo Kim, Jee Ye Kim, Young Kim, Suji Lee, Sol Moon, Ju-yong Hyon, Kyung-A Hyun, Yeji Yang, Seongmin Ha, Sunyoung Park, Hogyeong Gawk, Haeji Lee, Eun Hee Han, Jin Young Kim, Hyo-Il Jung, Young-Ho Chung, Seung Il Kim. Integrating machine learning with microfluidic technologies for proteomic profiling of extracellular vesicles in triple-negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1843.
Abstract 1845: 11,000-plex aptamer based proteomics of colorectal cancer stools for the discovery of novel disease biomarkers
Authors: Mohan Chandra , Vanarsa Kamala , Castillo Jessica , Bresalier Robert , Chia Nicholas
Published: March 2024

Given the morbidity and mortality associated with colorectal cancer, novel biomarkers are clearly warranted, especially for early detection, particularly for advanced adenomas. An earlier 1317-plex aptamer-based proteomic screen of stool proteins revealed 92 proteins to be significantly elevated in CRC stool, with the most discriminatory proteins being MMP9, haptoglobin, myeloperoxidase, fibrinogen, and adiponectin (PMID: 34117903). Results from a recent 2000-plex proteomic screen of CRC stools are detailed below, while a 11,000-plex aptamer based proteomic screen is in progress. Comprehensive proteomic screening revealed 116 proteins to be differentially expressed in CRC stool compared to control stool, with 45 being elevated 2-fold or higher. 37 of these proteins were ELISA validated in 3 independent cohorts. Stool MMP-8, MMP-9, Hemoglobin, PGRP-S, Haptoglobin, Myeloperoxidase and Fibrinogen emerged as being most discriminatory for distinguishing CRC from healthy controls (AUC: 0.91-0.95), while Stool Fibrinogen, MMP-9, Hemoglobin, MMP-8, and PGRP-S best distinguished advanced adenoma from healthy control (HC) stool, with stool Fibrinogen topping the list with an ROC AUC value of 0.86, as detailed in Table 1. Functional pathway analysis revealed a significant over-representation of pathways related to anti-oxidant activity, integrin/receptor binding, cytokines, blood coagulation, and lipoprotein biosynthesis in patients with CRC compared to HC. Nuclear Factor IC (NFIC) and IKZF2 were identified as key regulators of the molecular cascades over-represented in CRC. Stool Fibrinogen, MMP-8, MMP-9, PGRP-S, Haptoglobin, and Myeloperoxidase emerge as promising biomarkers for distinguishing CRC/advanced adenomas/healthy stools, outperforming Hemoglobin. Thus, comprehensive proteomics and independent ELISA validation have uncovered novel stool biomarkers for identifying CRC and advanced adenomas, that warrant further head to head comparison against current diagnostic tests.

Citation Format: Chandra Mohan, Kamala Vanarsa, Jessica Castillo, Robert Bresalier, Nicholas Chia. 11,000-plex aptamer based proteomics of colorectal cancer stools for the discovery of novel disease biomarkers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1845.
Abstract 1860: The smallest antibody class: Cow ultralong CDR3 knobs as a new class of biomolecule for therapeutics and diagnostics
Authors: Smider Vaughn
Published: March 2024

Engineered antibodies, including bispecifics and ADCs, are now a major drug class for oncology and other therapeutic areas. There are two fundamental antibody fragments: the Fv (heavy and light chain variable region fragments, derived from most species), and VHH (heavy chain ‘only’ V regions, derived from camelids and sharks). Cows produce antibodies with an unusually long VH CDR3 region, which can be up to seventy amino acids in length and is comprised of a β-ribbon “stalk” and disulfide-bonded “knob” minidomain which sits atop the stalk. The knob region is highly diverse and can bind cryptic epitopes with high affinity. We recently developed the technology to produce knob domains independently of the antibody, and at ~5 kDa are the smallest functional antibody fragment (https://doi.org/10.1073/pnas.2303455120). The high stability and small size could enable improved tissue penetration and development of novel therapeutics, including knob-drug conjugates or extremely small bispecific molecules.

Citation Format: Vaughn Smider. The smallest antibody class: Cow ultralong CDR3 knobs as a new class of biomolecule for therapeutics and diagnostics [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1860.
Abstract 1873: AST-05X: A novel GCN2 inhibitor as a promising anti-cancer approach for sarcoma
Authors: Choi Youngki , Lim Jinback , Park Hyo-Hyun , Kang Jinho , Kim Hyosong , Yun Kumhee , Jung Hun
Published: March 2024

Background Integrated stress response (ISR) is an adaptive signaling pathway that responds to stress and plays an important role in cell fate determination. This pathway enables cellular adaptation and survival under stress conditions by activating specific genes that reduce protein synthesis. Tumors exist in a state characterized by heightened intrinsic and extrinsic stress, heavily relying on a balanced ISR to manage the metabolic demands for rapid growth. ISR (Integrated Stress Response) consists of four major kinases: GCN2, PERK, HRI, and PKR. GCN2 is activated by stress due to amino acid deprivation. Amino acids are crucial for cell survival, and cancer cells, require higher amounts than normal cells, particularly depending on conditionally essential amino acids like glutamine, asparagine, and arginine. GCN2 activation acts as a key mechanism for cancer cells to overcome amino acid deficiencies, making GCN2 a potential target, especially in conjunction with anticancer drugs like asparaginase. In this study, we aimed to identify a potential GCN2 inhibitor to explore a promising anti-cancer approach in sarcoma, a rare cancer with no new therapeutic alternatives.

Method The activity verification of ISR kinases was characterized using in-vitro enzyme assays. In-vitro panel assays were employed to determine the selectivity profiling for oncology kinases. The modulation of the ISR pathway was validated using a sarcoma cell line (HT-1080) and a cell line derived from sarcoma patients. Western blot or ELISA assays were employed to measure phospho-GCN2, ATF4, phospho-eIF2a, and CHOP under both basal conditions and amino acid starvation conditions. Gene expression profiling (GEP) of sarcoma patient tissue samples(n=17) was conducted through RNA sequencing (RNAseq) analysis.

Results AST-05X is a selective and potent GCN2 inhibitor. AST-05X inhibits the expression of GCN2 and ISR downstream factors (p-eIF2a, ATF, CHOP) in vitro cell-based experiments under amino acid deprivation. In addition, based on the results of ISR-related gene expression profiling GEP analysis, we identified potential CDx biomarkers for evaluating the diagnosis and treatment response of sarcoma.

Conclusion This study is currently investigating the efficacy of the drug in sarcoma patient-derived xenograft models. AST-05X is a GCN2 inhibitor that has the potential to validate the inhibition of GCN2 as a promising anti-cancer approach for sarcoma, a rare cancer with limited therapeutic alternatives.

Citation Format: Youngki Choi, Jinback Lim, Hyo-Hyun Park, Jinho Kang, Hyosong Kim, Kumhee Yun, Hun Jung. AST-05X: A novel GCN2 inhibitor as a promising anti-cancer approach for sarcoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1873.
Abstract 1917: HER2-FGFR1 interaction induced by bispecific molecules: A novel strategy for treating breast cancer
Authors: Krzyscik Mateusz Adam , Hristova Kalina
Published: March 2024

Breast cancer poses a significant global health challenge, demanding effective therapies targeting specific receptors. In our prior study, we engineered molecules capable of binding to both HER2 and FGFR1, pivotal receptors in breast cancer progression. By combining the high-affinity Affibody ZHER2:2891 for HER2 with a modified, stable form of FGF2 binding to FGFR1, we created the bispecific protein (AfHER2-lFGF2). This innovative molecule could simultaneously recognize HER2 and FGFR1, exhibiting enhanced cell penetration. When coupled with the potent cytotoxic agent monomethyl auristatin E (MMAE), it demonstrated exceptional efficacy in eliminating cancer cells expressing HER2, FGFR, or both receptors. Our current investigation focused on quantifying the interaction between HER2 and FGFR1 receptors on the cell surface upon treatment with these bispecific molecules. Leveraging fully quantified spectral imaging FRET (FSI-FRET), a technique utilizing spectrally resolved two-photon microscopy. We directly monitor the association of HER2 and FGFR1 in the plasma membrane. Our findings revealed that the bispecific AfHER2-lFGF2 molecule forces interaction between HER2 and FGFR1 on the cell surface. Furthermore, we analyzed the activation of the HER2-FGFR1 complex and subsequent downstream signaling pathways using western blotting. In summary, our research uncovers the ability of the AfHER2-lFGF2 bispecific protein to foster direct interaction between HER2 and FGFR receptors on the cell membrane. By employing state-of-the-art techniques like FSI-FRET, we elucidated the mechanisms governing this interaction and, by western blotting, downstream signaling pathways. These insights emphasize the potential of AfHER2-lFGF2 as a promising therapeutic agent for targeting HER2, FGFRs, and HER2/FGFRs-positive cancers. This approach paves the way for targeted therapies in addressing the prevalent global health challenge of breast cancer.

Citation Format: Mateusz Adam Krzyscik, Kalina Hristova. HER2-FGFR1 interaction induced by bispecific molecules: A novel strategy for treating breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1917.
Abstract 1928: Targeting FAK to improve the therapy of KRAS G12C mutant cancers
Authors: Liu Shuang , Shi Junfen , Lu Chang , Zhang Miao , Xin Ying , Zhu Yan , Li Jijun , Luo Hong , Sun Yinghui
Published: March 2024

Despite the pivotal role of KRAS G12C in tumor initiation and progression, currently available KRAS G12C inhibitors have relatively modest activity compared to other approved therapies targeting other classic oncogenic drivers. Fortunately, efforts to improve the anti-tumor response to KRAS G12C targeted therapy have benefited from the use of combination approaches. Here, we found that treatment with KRAS G12C inhibitors induces sustained activation of focal adhesion kinase (FAK) in KRAS G12C mutant cell lines from various cancer types, including non-small cell lung cancer, colorectal cancer, and pancreatic cancer. These results suggest a potential role for FAK in adaptive resistance to KRAS G12C inhibition and encourage us to explore the anticancer effects of combining the KRAS G12C inhibitor SY-5933 with a FAK inhibitor, CT-707. The cell viability and clonogenic assays using various KRAS G12C mutant cancer cell lines showed that the combination of SY-5933 and CT-707 synergistically inhibited cell growth. Mechanistically, this enhanced anti-proliferative effect is associated with a significant decrease in FAK activity and a more pronounced induction of cell cycle arrest and apoptosis. In vivo efficacy studies in multiple cell line derived xenograft (CDX) models, the combination of SY-5933 plus CT-707 consistently induced a more potent anti-tumor response than either monotherapy alone. In conclusion, our results demonstrate that CT-707 has the potential to enhance the efficacy of SY-5933 by suppressing adaptive FAK activation under KRAS G12C inhibition. This provides a rationale for an innovative combination therapy to improve outcomes in patients with KRAS G12C mutated cancers.

Citation Format: Shuang Liu, Junfen Shi, Chang Lu, Miao Zhang, Ying Xin, Yan Zhu, Jijun Li, Hong Luo, Yinghui Sun. Targeting FAK to improve the therapy of KRAS G12C mutant cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1928.
Abstract 1948: Resistance to RAS inhibition promotes remodeling of the immunosuppressive microenvironment of pancreatic cancer
Authors: Tiriac Herve
Published: March 2024

Background: Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal malignancies, largely due to acquired chemotherapy resistance. MRTX-1133, a promising KRAS-G12D inhibitor, has demonstrated initial efficacy; however, resistance analogous to that seen with KRAS-G12C inhibitors is an emerging challenge. The tumor microenvironment (TME) is increasingly recognized as a contributor to resistance, often fostering tumor survival and immune evasion.

Methods: To elucidate the resistance mechanisms to KRAS-G12D inhibition, we established resistant mouse and human PDAC cell lines derived from mouse and human models. These models were analyzed using flow cytometry and single cell RNA sequencing to define TME dynamics as resistant is acquired and after is has been established. We also analyzed cytokines released by resistant tumor cells using protein arrays and RNA sequencing.

Results: Our investigation revealed significant TME alterations associated with acquired resistance. Notably, there was a marked increase in the population CD4+ Tregs and neutrophil while activated CD8+ T cells are decreased in the resistant tumors. Concurrently, an increased secretion of chemokines such as CXCL1 and CXCL2 was observed in resistant tumor cells relative to their parental counterparts.

Conclusions: The adaptation of PDAC to circumvent KRAS inhibition involves a distinct reprogramming of the TME, attenuating immune surveillance and potentially facilitating tumor progression. Current efforts are directed towards delineating the key pathways implicated in these TME modifications to pioneer new therapeutic approaches for managing KRAS inhibitor resistant PDAC.

Citation Format: Herve Tiriac. Resistance to RAS inhibition promotes remodeling of the immunosuppressive microenvironment of pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1948.
Abstract 1974: Discovery of TRX-211-399, a potent, CNS-penetrant, highly selective inhibitor of EGFR exon 20 insertion mutations
Authors: Chun Kwangwoo , Lim Sumin , Lee Joohyun , Choi Seulgi , Kim Seongrak , Choi Youngil , Park Areum , Kim Bohee , Park Yongseo , Choi Eunhyun , Lee Sungwon , Lee Koo
Published: March 2024

Lung cancer is the leading cause of cancer-related deaths worldwide, accounting for approximately 2 million new cases and 1.8 million deaths annually. Various types of EGFR-activating mutations are one of the most important oncogenic drivers in non-small cell lung cancer (NSCLC). Among NSCLC patients with EGFR mutations, 5-10% of patients have EGFR exon 20 insertion (ex20ins) mutations that are insensitive to clinically available EGFR tyrosine kinase inhibitors (TKIs). Recently, as mobocertinib (EXKIVITY®) failed to improve progression-free survival in Phase 3 clinical trial, there remains a high unmet medical need to develop new drugs with wider therapeutic window to treat NSCLC harboring EGFR ex20ins mutations. Herein, we report a novel EGFR-TKI, TRX-211-399, specifically designed to target ex20ins mutations. TRX-211-399 potently inhibited the proliferation of not only Ba/F3 cells overexpressing near-loop (A767dupASV, S768dupSVD) and far-loop (H773insNPH, H773dupH) mutations but also patient-derived NSCLC cell line. In particular, TRX-211-399 demonstrated high selectivity against wild-type EGFR. With favorable DMPK profiles, once-daily oral administration of TRX-211-399 showed significant tumor regression in multiple Ba/F3 xenograft models carrying different EGFR exon20ins mutations. Furthermore, in a PC-9-luc cell intracranial xenograft model, TRX-211-399 induced substantial anti-tumor effects and prolonged survival without body weight loss. TRX-211-399 is currently undergoing further preclinical evaluation as a potential candidate for clinical development for patients with EGFR exon20ins mutations.

Citation Format: Kwangwoo Chun, Sumin Lim, Joohyun Lee, Seulgi Choi, Seongrak Kim, Youngil Choi, Areum Park, Bohee Kim, Yongseo Park, Eunhyun Choi, Sungwon Lee, Koo Lee. Discovery of TRX-211-399, a potent, CNS-penetrant, highly selective inhibitor of EGFR exon 20 insertion mutations [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1974.
Abstract 1993: Cytosolic BAX dimerization as a mechanism of resistance to apoptosis and a novel target to potentiate BAX activation and apoptosis
Authors: Gitego Nadege , Agianian Bogos , Mak Oi Wei , MV Vasantha Kumar , Cheng Emily , Gavathiotis Evripidis
Published: March 2024

BCL-2 family proteins are master regulators of cellular life and death and have become drug targets for the development of therapeutic agents to overcome the apoptotic blockade of cancer. BAX is a pro-apoptotic BCL-2 member that is a crucial mediator of apoptosis induced by diverse stimuli including various chemotherapeutic and targeted agents. BAX predominantly resides in the cytosol in a quiescent state and upon stress, it undergoes conformational activation and mitochondrial translocation leading to mitochondrial outer membrane permeabilization, a critical event in apoptosis execution. While cytosolic BAX is typically considered an inactive monomer, we previously reported structural and cell-based evidence of a cytosolic BAX dimer. However, it remains unclear how this regulates BAX. Moreover, we found that specific BAX mutants such as P168G and G67R formed inactive BAX dimers and mutations of P168A and G67R have been identified in AML patients and found to have impaired apoptotic activity and notably the BAX P168A mutant appeared in AML patients acquiring resistance to the BCL-2 inhibitor Venetoclax. Here, we investigated the role of the inactive BAX dimer in regulating apoptosis and pro-apoptotic drug treatments in various hematologic and solid tumor cancer cells. Surprisingly, cancer cell lines express cytosolic inactive BAX dimers and/or monomers. Expression of inactive dimers, results in reduced BAX activation, translocation and apoptosis upon pro-apoptotic drug treatments such as BH3 mimetics and chemotherapy drugs. Our findings support the notion that formation of the inactive BAX dimer is a mechanism adopted by cancer cells to further suppress BAX activation and gain survival advantage. Using the inactive BAX dimer crystal structure, we designed a pharmacophore-based drug screen, and identified a small-molecule modulator, BDM19 that binds and activates cytosolic BAX dimers and prompts cells to apoptosis alone or in combination with BCL-2/BCL-XL inhibitors. Moreover, we validated interactions of BDM19 and BAX, using structural, biochemical and cellular methods, critical for BAX activation. Our findings provide mechanistic insights into the regulation of BAX and resistance to apoptosis in cancer through inactive BAX dimerization. Further, we demonstrate a rational strategy to target BAX dimer for therapeutic modulation of BAX and overcoming resistance to apoptosis by pro-apoptotic agents.

Citation Format: Nadege Gitego, Bogos Agianian, Oi Wei Mak, Vasantha Kumar MV, Emily Cheng, Evripidis Gavathiotis. Cytosolic BAX dimerization as a mechanism of resistance to apoptosis and a novel target to potentiate BAX activation and apoptosis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1993.
Abstract 1994: Drug-induced cancer cell secretome promotes resistance in colon cancer
Authors: Satapathy Shakti Ranjan , Ghatak Souvik , Sjölander Anita
Published: March 2024

Colorectal cancer (CRC) is the third most common cancer in the world, with a projection of increased lethality in the upcoming years. Adjuvant chemotherapy with biological agents is the most practiced standard of care today. However, patients with metastatic CRC (mCRC) do not benefit from the wide range of therapies available for CRC, which either owe to unresectability or co-morbidity or more importantly resistance to therapy. 5-fluorouracil (5-FU) combined with leucovorin, oxaliplatin, or irinotecan (FOLFOX or FOLFIRI) has been routinely employed as the first-line therapy in colon cancer, but patients often develop resistance to 5-FU. Additionally, tumor cell dormancy plays a major role in tumor relapse which leads to poor prognosis in the patients. Despite many advances, the understanding of therapy-induced resistance is still in its infancy. Here, we show that 5-FU-induced tumor cell secretome helps in the outgrowth and metastasis of 5-FU-resistant clones in colon cancer. We used parental and 5-FU-resistant colon cancer cell lines, and transgenic zebrafish embryos to test the hypothesis and perform the colon cancer cell-based functional assays. We observed that 5-FU resistant (5-FU-R) colon cancer cells showed an increase in proliferation and survival when cultured with the secretome of 5-FU exposed colon cancer cells compared to the DMSO-induced secretome. Interestingly, we also noted elevated migration and invasion in the 5-FU-R colon cancer cells exposed to 5-FU-induced secretome compared to the DMSO counterpart. Additionally, in zebrafish xenograft, we found that 5-FU-R colon cancer cells cultured with 5-FU-induced secretome showed higher tail-vein metastatic burden compared to the 5-FU-R cells cultured with DMSO-induced secretome. This provides a background to further study in detail the therapy-induced resistance as well as dormancy in colorectal cancer which will help to develop approaches to prevent or reverse chemoresistance in patients who receive systemic therapy for mCRC.

Citation Format: Shakti Ranjan Satapathy, Souvik Ghatak, Anita Sjölander. Drug-induced cancer cell secretome promotes resistance in colon cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1994.
Abstract 2020: Determination of parameters to understand the metabolism in the tumor microenvironment by in vivo microdialysis
Authors: Heins Mariette Suzan , Flik Gunnar
Published: March 2024

The tumor microenvironment presents a unique surrounding with more favorable conditions for the cancer cells to growth thrive and interact than for the host cells. To obtain a better understanding of cancer biology and pathophysiology, information about the physiological state of the tumor microenvironment is crucial. By using the in situ microdialysis technique we can quantitate relevant modulators in the interstitial space of the tumor.

The current set of experiments demonstrates the benefits of the use of microdialysis in freely-moving awake tumor-bearing animals. Following inoculation, growth was monitored continuously and upon an average tumor volume of 400 mm3, the animals underwent surgical implantation of microdialysis probes. The next day, the sample collection was initiated by perfusion of the implanted probes with dialysate fluid. Samples were continuously collected over several hours and stored at -80C. Levels of analytes indicative of metabolic and physiological conditions of the tumor microdialysis were quantified by EIA and/or LC-MS/MS analysis.

The microdialysis technology combined with sensitive analytical methods allows us to quantify several analytes from the tumor microenvironment simultaneously and monitor changes over time. Metabolic activity is an important indicator of tumor growth speed. Depletion of energy can lead to increases in lactate levels and acidification of the tumor microenvironment. Resulting in a reduction of host immunological efficacy. Elucidation of the status of the tumor environment during tumor growth and treatment allows for better translational models.

Citation Format: Mariette Suzan Heins, Gunnar Flik. Determination of parameters to understand the metabolism in the tumor microenvironment by in vivo microdialysis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2020.
Abstract 204: Quantitative evaluation of biomarkers for TGF-β-induced epithelial-mesenchymal transition in biochemical, cellular, and 3D spheroid model systems
Authors: Wood Antony W. , Heimsath Ernest
Published: March 2024

Epithelial-mesenchymal transition (EMT) is a cellular differentiation process whereby epithelial cells lose epithelial features and acquire mesenchymal, fibroblast-like properties, leading to reduced cell-cell contacts and increased motility. EMT is a fundamental biological process required for normal embryonic development, but may be co-opted by malignant epithelial tumors to facilitate metastatic spread. A known driver of EMT is the TGF-β superfamily of growth factor ligands, which elicit receptor-mediated responses in cells, primarily via TGF-β/SMAD signaling pathways. In these pathways, receptor-mediated SMAD (R-SMAD) proteins serve as the primary downstream effector molecules, the activities of which are regulated through receptor-mediated phosphorylation. The magnitude and duration of ligand-induced receptor activation influences the level of SMAD phosphorylation, which in turn influences the magnitude of the downstream cellular response(s). In this study, we describe high-throughput methods to quantitatively evaluate the biochemical and cellular responses to TGF-β/SMAD pathway activation in a cellular model of TGF-β-induced EMT. Effects of pathway activation are examined at different levels of biological complexity (biochemical, cellular, and multicellular), using 2D and 3D (spheroid) models. Collectively, these approaches enable a comprehensive evaluation of TGF-β/SMAD pathway activation that is amenable to high-throughput analysis platforms.

Citation Format: Antony W. Wood, Ernest Heimsath. Quantitative evaluation of biomarkers for TGF-β-induced epithelial-mesenchymal transition in biochemical, cellular, and 3D spheroid model systems [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 204.
Abstract 2049: Novel preclinical models for PROTAC evaluation: And beyond
Authors: xu yuzhou , Zhang Jichuan , Tian Ye , Chen Ziyu , Wang Amy , Ma Ting , Chen Qikuan , Yin Yinfei
Published: March 2024

Proteolysis-targeting chimeras (PROTACs) are a novel class of small molecules that induce the degradation of target proteins. Unlike conventional inhibitors that disable target proteins by direct binding, PROTACs are event-driven, meaning that a single PROTAC molecule can induce the degradation of multiple target protein molecules. This catalytic mechanism allows PROTACs to achieve complete inhibition of target proteins at lower concentrations compared to conventional inhibitors, making them a promising therapeutic approach for previously considered "undruggable" targets.Despite their promise, the development of PROTACs faces several challenges. The large molecular weight of PROTACs can lead to poor cell permeability and bioavailability. Additionally, the lack of high-throughput, quantitative assays for evaluating PROTAC efficacy hinders structure-activity relationship (SAR) studies. The formation of the Protein-PROTAC-E3 ternary complex is crucial for PROTAC function, and reliable methods for assessing this interaction are essential for PROTAC development.In this poster, ChemPartner showcases a comprehensive suite of preclinical models for PROTAC evaluation, encompassing both in vitro and in vivo approaches. Using low-cost, high-throughput biochemical assays, we can provide valuable insights into PROTAC efficacy, facilitating SAR studies and elucidating the relationship between molecular structure and activity. Followed by in cell and in vivo function assays, we can test PROTAC PK/PD, toxicity, and efficacy. This suite of assays provides a powerful tool to speed up PROTAC development and reduce the fail rate.

Citation Format: yuzhou xu, Jichuan Zhang, Ye Tian, Ziyu Chen, Amy Wang, Ting Ma, Qikuan Chen, Yinfei Yin. Novel preclinical models for PROTAC evaluation: And beyond [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2049.
Abstract 2090: The GNAQ T96S mutation enhances the oncogenic properties of breast cancer
Authors: Wu Ming-Hsi , Wang Hsiang-Ching , Wang Wei-Hsin , Lin Yen-Ju , Chen Wan-Ru , Chen Shih-Ta , Fu Chih-Wei , Chuang Eric
Published: March 2024

Breast cancer (BC) is the most commonly diagnosed cancer in the world, and metastatic breast cancer (mBC) remains responsible for the majority of breast cancer deaths. In this study, we collected 615 cases of Taiwanese BC patient's tumor samples and used whole exome sequencing (WES) to clarify the genetic mutation landscape of this population. We have compared with various BC genomic databases such as TCGA, COSMIC, and MSKCC to identify 29 novel variants in 18 genes that were predicted to have a pathogenic effect. In these pathogenic variants, we found that the GNAQ T96S mutation (threonine 96 to serine alteration of the Gαq protein) was presented in 116 out of 615 BC patients (18.9%). To examine the effect of the GNAQ T96S mutation on BC, we transfected the MCF-7 and MDA-MB-231 cell lines with the wild-type or the mutant GNAQ T96S expression vector. Transfection with the GNAQ T96S expression vector enhanced cell proliferation, colony formation, migration, and activation of the MAPK pathways compared to wild-type GNAQ expression MCF-7 cells. We also found that the tumorigenicity, and tumor growth were significantly increased in the GNAQ T96S expressed MCF-7 xenograft model, and exhibited higher rates of lung metastasis in GNAQ T96S expressed MDA-MB-231 cells. Our data suggest that the GNAQ T96S mutation may play an oncogenic role in BC by potentiating the GNAQ signaling pathway. In this study, we also found several compounds that can inhibit in vitro cell proliferation and in vivo tumor growth of GNAQ T96S expressed MCF-7 cells. These compounds showed the potential to treat the mBC patients who have GNAQ T96S mutation.

Citation Format: Ming-Hsi Wu, Hsiang-Ching Wang, Wei-Hsin Wang, Yen-Ju Lin, Wan-Ru Chen, Shih-Ta Chen, Chih-Wei Fu, Eric Chuang. The GNAQ T96S mutation enhances the oncogenic properties of breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2090.
Abstract 2153: The impact of socioeconomic and racial segregation on cancer stage at diagnosis, treatment and mortality among patients with small cell lung cancer
Authors: Bekele Bayu B. , Lian Min , Liu Ying
Published: March 2024

Background: Although studies have revealed the adverse impact of residential segregation on early detection, treatment, and survival of non-small cell lung cancer―a predominant type of lung cancers, it remains unknown regarding the role of residential segregation in treatment and outcomes of small cell lung cancer (SCLC)―a rare type of lung cancer characterized by rapid growth and poor prognosis. Therefore, we examined the associations of racialized socioeconomic segregation with risks of extensive-stage diagnosis, non-adherence to guideline-recommended treatment, and mortality among non-Hispanic White (NHW) and non-Hispanic Black (NHB) patients with SCLC.

Methods: This population-based retrospective cohort study included patients who were diagnosed with SCLC as a first primary malignancy at age 20 and older between January 2007 and December 2015, followed through December 2016, and identified from the Surveillance, Epidemiology, and End Results. County-level racialized socioeconomic segregation was measured using the Index of Concentration at the Extremes (ICE). Multilevel logistic regression accounting for county-level clustering was used to estimate odds ratios (ORs) for extensive-stage diagnosis and non-adherence to guideline-recommended treatment. Hazard ratios (HRs) for lung cancer-specific mortality and all-cause mortality were computed using multilevel Cox regression.

Results: Among 38,393 patients, 90.1% were NHW and 9.9% were NHB. Patients dwelling in the counties with the highest concentration of low-income NHB households (highest ICE quartile) had higher risks of non-adherence to guideline-recommended treatment (OR=1.23, 95% CI: 1.08-1.40; Ptrend <0.01), lung cancer-specific mortality (HR=1.08, 95% CI: 1.04-1.12; Ptrend <0.01), and all-cause mortality (HR=1.13, 95% CI: 1.09-1.17; Ptrend <0.0001) as compared with patients dwelling in the counties with the highest concentration of high-income NHW households (lowest ICE quartile). These associations were comparable between NHB and NHW patients. Racialized economic segregation was not significantly associated with extensive-stage diagnosis.

Conclusions: Residing in the segregated, low-income NHB counties had adverse impacts on stage-appropriate treatment for SCLC and survival outcomes. Future research on the pathway(s) linking residential segregation to SCLC treatment underutilization and poor outcomes is warranted.

Citation Format: Bayu B. Bekele, Min Lian, Ying Liu. The impact of socioeconomic and racial segregation on cancer stage at diagnosis, treatment and mortality among patients with small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2153.
Abstract 2182: 15-lipoxygenase-1 modulation of colonic resolvin production to suppress colorectal carcinogenesis
Authors: Zuo Xiangsheng , Kiyasu Yoshiyuki , Liu Yi , Moussalli Micheline , Wei Bo , Yang Peiying , Shureiqi Imad
Published: March 2024

Docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) are widely used dietary supplements and FDA-approved treatments. The reported impacts of fish oil, DHA, and EPA on colorectal carcinogenesis (CRC) have spurred debates, as certain studies suggest promotional effects while numerous others emphasize suppressive effects. Nevertheless, resolvins, oxidative metabolites of EPA and DHA, exhibit the ability to inhibit crucial pathways (such as TNF-α and IL-1β) that contribute to CRC. 15-lipoxygenase-1 (ALOX15) plays a crucial role in the oxidative metabolism of DHA and EPA, ultimately leading to the formation of resolvins. However, ALOX15 expression is commonly lost during human CRC. Whether loss of ALOX15 expression modulates the effects of DHA and EPA on CRC remains to be elucidated. We therefore conducted the experiments in preclinical CRC mouse models induced by Azoxymethane or Apc gene mutation to explore the hypothesis that ALOX15 expression in colonic epithelial cells regulates DHA and EPA metabolism to generate resolvins, subsequently influencing CRC outcomes. We found that 1) DHA and EPA exert differential effects on CRC, which can be further modified by factors such as their formulation (e.g., ethyl ester versus triglyceride); 2) targeted transgenic expression of ALOX15 in the intestines of mice (referred to as ALOX15-Gut mice) markedly augmented the production of resolvins and concurrently suppressed CRC; 3) the predominant products from the expressed ALOX15 enzymatic activity were identified as 17-HDHA and resolvin D5; and 4) the effects ALOX15 on increasing resolvin generation and suppressing CRC were consistent across various promoters used to drive ALOX15 transgenic expression in mice. These findings collectively underscore the profound importance of ALOX15 intestinal expression as a host-related factor in resolvin generation from EPA and DHA, which subsequently contributes to the suppression of CRC.

Citation Format: Xiangsheng Zuo, Yoshiyuki Kiyasu, Yi Liu, Micheline Moussalli, Bo Wei, Peiying Yang, Imad Shureiqi. 15-lipoxygenase-1 modulation of colonic resolvin production to suppress colorectal carcinogenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2182.
Abstract 2194: Association of metabolic syndrome conditions with risk of second primary cancer of any organ in breast cancer survivors
Authors: Mukherjee Amrita , Gu Zheng , Chen Lie Hong , Potosky Arnold L. , Haque Reina
Published: March 2024

Purpose: Breast cancer survivors are at increased risk of developing second primary cancers; however, factors attributing to increased risk of second primary cancer in breast cancer survivors are under-studied. We evaluated the association of metabolic syndrome conditions with second primary cancer of any organ in breast cancer survivors.

Methods: In this retrospective cohort study, we included 37,222 patients diagnosed with breast cancer between 2008-2020 at Kaiser Permanente Southern California (KPSC), an integrated healthcare system. We extracted data on cancer-related variables, sociodemographic and clinical variables from KPSC’s cancer registry and electronic health records. Among the metabolic conditions, we identified diabetes, hypertension, and dyslipidemia using ICD-10 diagnosis codes at or 1-year prior to breast cancer diagnosis; obesity was based on body mass index (BMI≥30kg/m2). We included metabolic syndrome as a categorical variable: 0, 1-2, and 3+ metabolic conditions. We followed patients electronically from breast cancer diagnosis until patients developed second primary cancer of any organ, died, disenrolled from the health plan, or reached study’s end (12/31/2021), whichever occurred first. Proportional hazards regression was used to report association [HR (95%CI)] between metabolic conditions and any second primary cancer.

Results: During a median (IQR) follow-up of 5.17 (2.75-8.58) years, 1701 (4.6%) patients developed any second primary cancer. More than half (53.1%) of the breast cancer survivors had 1-2 metabolic conditions at baseline; 19.4% had 3+ metabolic conditions, and the rest 27.5% did not have any metabolic condition. Hypertension was the most common metabolic condition (45.5%), followed by obesity (38.9%), dyslipidemia (36.5%), and diabetes (19.0%). Survivors with 0 and 1-2 metabolic conditions were 35% and 19% less likely to develop any second primary cancer than survivors with 3+ metabolic conditions, respectively [crude HR (95%CI): 0.65 (0.57-0.75) and 0.81 (0.72-0.91), respectively]. Reduced risk in survivors with 1-2 metabolic conditions persisted adjusting for age, race and ethnicity, breast cancer diagnosis year, breast tumor SEER summary stage, breast tumor estrogen-progesterone status, smoking status, and breast tumor treatment [adjusted HR (95%CI): 0.87 (0.77-0.99)].

Conclusion: Risk of second primary cancer of any organ was elevated in breast cancer survivors with three or more metabolic conditions.

Citation Format: Amrita Mukherjee, Zheng Gu, Lie Hong Chen, Arnold L. Potosky, Reina Haque. Association of metabolic syndrome conditions with risk of second primary cancer of any organ in breast cancer survivors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2194.