bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1195: Prospective study for the identification of nivolumab biomarkers via analyses of pre-treatment plasma exosome mRNAs from head and neck cancer patients (BIONEXT)
Authors: Masuda Muneyuki , Toh Satoshi , Matsuo Mioko , Sugasawa Masashi , Yamazaki Keisuke , Yushi Ueki , Nakashima Torahiko , Uryu Hideoki , Ono Takeharu , Ueda Tsutomu , Umeno Hirohito , Kano Satoshi , Tsukahara Kiyoaki
Published: March 2024

Nivolumab paved a new way in the treatment of recurrent or metastatic (RM) head and neck squamous cell carcinoma of (HNSCC). However, the limited rates of long-term survivors (< 20%) demand a robust prognostic biomarker. This nationwide multi-centric prospective study aimed to identify a plasma exosome (PEX) mRNA signature, which serves as a companion diagnostic of nivolumab and as well provides a biological clue to develop effective therapies for a majority of non-survivors. Pre-treatment plasmas (N = 104) were subjected to comprehensive PEX mRNA analyses for prognostic marker discovery and validation. In parallel, paired treatment naïve tumor and plasma samples (N = 20) were assayed to elucidate biological implications of the PEX mRNA signature. A combination of 6 candidate PEX mRNAs plus neutrophil-to-lymphocyte ratio precisely (P = 0.000124) distinguished non-survivors from >2-yr survivor (0% vs 57.7%) with a high hazard ratio (2.878; 95% confidence interval 1.639-5.055; P = 0.0002348). In paired samples, high PEX HLA-E mRNA (a non-survivor-predicting marker) reflected the increased HLA-E protein expression (P = 0.0191) and the dense population of tumor-infiltrating NK cells (P = 0.024) in the corresponding tumor. It is estimated that the effects of PD-1blockade is canceled by the HLA-E-NKG2A immune checkpoint in patients with high PEX HLA-E mRNA. The combination of NKG2A antibody (i.e., monalizumab) and nivolumab may be a promising strategy for non-survivors predicted by a RT-qPCR-based pre-treatment measurement of PEX mRNAs. A candidate companion diagnostic of nivolumab that indicates a possible treatment option is identified.

Citation Format: Muneyuki Masuda, Satoshi Toh, Mioko Matsuo, Masashi Sugasawa, Keisuke Yamazaki, Ueki Yushi, Torahiko Nakashima, Hideoki Uryu, Takeharu Ono, Tsutomu Ueda, Hirohito Umeno, Satoshi Kano, Kiyoaki Tsukahara. Prospective study for the identification of nivolumab biomarkers via analyses of pre-treatment plasma exosome mRNAs from head and neck cancer patients (BIONEXT) [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 1195.
Abstract 1205: Inferring allele-specific copy number aberrations and tumor phylogeography from spatially resolved transcriptomics
Authors: Ma Cong , Balaban Metin , Liu Jingxian , Chen Siqi , Ding Li , Raphael Benjamin J.
Published: March 2024

Tumors evolve through the acquisition of somatic mutations over time and across space. While the temporal aspects of tumor evolution can be inferred by applying phylogenetic techniques to bulk or single-cell DNA sequencing data, the spatial aspects of tumor evolution remain understudied. Spatially resolved transcriptomics (SRT) measures gene expression at thousands of spatial locations in a tumor, but does not directly measure genetic aberrations. Thus, SRT data has not yet been fully utilized for tumor evolution studies. We introduce CalicoST, an algorithm that simultaneously infers allele-specific copy number aberrations (CNAs) and derives a phylogeographic model of tumor evolution from SRT data. By modeling CNA-induced perturbations in both gene expression and allele frequencies, CalicoST identifies important types of CNAs - including copy-neutral loss of heterozygosity (CNLOH) and mirrored subclonal CNAs - that are invisible to total copy number analysis. On SRT data from nine patients from the Human Tumor Atlas Network (HTAN), CalicoST achieves an average accuracy of 86%, approximately 21% higher than existing methods. On two of these patients with multiple adjacent slices, CalicoST reconstructs a tumor phylogeography that describes the spread of cancerous clones in three-dimensional space. CalicoST analysis of multiple SRT slices from a cancerous prostate organ reveals five spatially coherent clones, with mirrored subclonal CNAs distinguishing clones on the two sides of the prostate, and a bifurcating phylogeography in both genetic and physical space that suggests a polyclonal cancer.

Citation Format: Cong Ma, Metin Balaban, Jingxian Liu, Siqi Chen, Li Ding, Benjamin J. Raphael. Inferring allele-specific copy number aberrations and tumor phylogeography from spatially resolved transcriptomics [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 1205.
Abstract 1216: Pan-RAS inhibition by a tumor-targeted biotherapeutic
Authors: Beilhartz Greg L. , Liang Huazhu , Udaskin Molly , Ng Christine , Radulovich Nikolina , Melnyk Roman A.
Published: March 2024

Oncogenic RAS signaling drives tumor growth in about 30% of human malignancies. In recent years the first mutant-specific inhibitors of KRAS G12C (sotorasib and adagrasib) entered the clinic. While some durable responses were noted in KRAS-mutant non-small cell lung cancer, the clinical experience of KRAS G12C inhibitors is marked by rapid drug resistance, either intrinsic or acquired, causing re-activation of the MAPK pathway. Common resistance mechanisms include compensatory signaling through the related RAS isoforms NRAS and HRAS, as well as the selection of non-G12C KRAS mutations, among others. The bacterial peptidase RRSP potently cleaves all RAS mutants and isoforms in the conserved Switch I region. We have shown that receptor-mediated intracellular delivery of RRSP using an engineered protein delivery system (RASx) can inactivate all RAS signaling in a cell, causing apoptosis in RAS-addicted tumor cells in vitro and in vivo. Since systemic pan-RAS inhibition is not likely to be tolerated therapeutically, precise tumor targeting is a necessary feature of a true pan-RAS inhibitor. Here, we describe the unique ability of the RASx system to specify the cell surface receptor used by RRSP to enter cells, allowing exquisite targeting and killing of KRAS, NRAS and HRAS-driven tumor cells while sparing healthy tissues. We screened a panel of patient-derived tumor organoids (PDOs), to demonstrate broad tumor cell killing across pancreatic and colorectal cancer-derived PDOs. 97% of KRAS mutant organoids were sensitive to RASx, independent of the specific mutation. Intriguingly, 46% of wildtype RAS PDOs were sensitive to RASx, including organoids with activating mutations in EGFR. In contrast, PDOs with mutations downstream of RAS such as BRAF V600E were resistant to RASx. Indeed, RASx is effective not only against RAS-addicted tumors, but also wildtype RAS tumors driven by upstream receptor tyrosine kinases, potentially expanding the clinical utility of this platform. RASx is a first-in-class, tumor-targeted pan-RAS inhibitor that represents a unique entry into the growing armamentarium against oncogenic RAS.

Citation Format: Greg L. Beilhartz, Huazhu Liang, Molly Udaskin, Christine Ng, Nikolina Radulovich, Roman A. Melnyk. Pan-RAS inhibition by a tumor-targeted biotherapeutic [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 1216.
Abstract 1242: Foxa1/2-dependent epigenomic reprogramming drives lineage switching in lung adenocarcinoma
Authors: Gillis Katherine , Orellana Walter , Wilson Emily , Fort Gabriela , Dadzie Headtlove Essel , Parnell Timothy , Zhang Xiaoyang , Snyder Eric
Published: March 2024

The ability of cancer cells to alter their identity is essential for tumor survival and progression. Loss of the pulmonary lineage specifier NKX2-1 within KRAS-driven lung adenocarcinoma (LUAD) enhances tumor progression and results in a pulmonary-to-gastric lineage switch that is dependent upon the activity of pioneer factors FoxA1 and FoxA2; however, the underlying mechanism remains largely unknown. Here, we perform transcriptional and epigenetic assays to show that FoxA1/2 reprogram the chromatin landscape of NKX2-1-negative LUAD to facilitate a gastric differentiation program. Using sequential recombination models, we find that FoxA1/2 are required for demethylation of gastric-defining genes after Nkx2-1 deletion. FoxA1 colocalizes with TET3, an enzyme that mediates DNA demethylation, in NKX2-1-negative tumors. Deletion of Foxa1/2 results in loss of TET3 occupancy at key gastric marker genes, indicating that FoxA1/2 recruit TET3 to lineage-specific sites. H3K27ac ChIP-seq and HiChIP show that FoxA1/2 also control the activity of enhancers and promoters as well as their 3D interactions at gastric target genes following NKX2-1 loss. Furthermore, oncogenic KRAS is required for the FoxA1/2-dependent epigenetic reprogramming in NKX2-1-negative LUAD. This work demonstrates the role of FoxA1/2 in rewiring the methylation and histone landscape and cis-regulatory dynamics of NKX2-1-negative LUAD to drive cancer cell lineage switching.

Citation Format: Katherine Gillis, Walter Orellana, Emily Wilson, Gabriela Fort, Headtlove Essel Dadzie, Timothy Parnell, Xiaoyang Zhang, Eric Snyder. Foxa1/2-dependent epigenomic reprogramming drives lineage switching in lung adenocarcinoma [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 1242.
Abstract 1297: Patient engagement: Improving the at home care for cancer patients in treatment
Authors: Blomquist Robert
Published: March 2024

CareBOX Program is an Austin-based non-profit founded in 2014 to address a serious gap in cancer care for patients in the Central Texas area. It provides essential nutritional supplements and supplies to help prevent malnutrition, infections, and injuries from falls as patients continue at-home recovery during treatment and following surgery. About 80 percent of the supplies CareBOX Program provides are not covered by insurance. Many patients could not afford to purchase these items on their own. Volunteers deliver the CareBOXes to the clients’ homes. CareBOX Program has always been an inclusive organization. We operate on the basis that all cancer patients in Central Texas in need of our services will receive them as long as we have the funding to do so. Our youngest patient has been 3 months old and our oldest patient has been 93 years old. Currently we are serving more women (61%) than men (39%). Approximately 67% of patient households report annual income of less than $35,000. To date, CareBOX program volunteers have delivered more than 3,800 CareBOXes

Citation Format: Robert Blomquist. Patient engagement: Improving the at home care for cancer patients in treatment [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 1297.
Abstract 1314: SCHEQ Foundation (STEMM & Cancer Health Equity): Increasing STEMM diversity and outcomes for patients navigating cancer
Authors: Manley Eugene
Published: March 2024

Health inequities are estimated to cost $320B annually, while racial and ethnic disparities are estimated to cost $42B in productivity. There are systemic issues at play that contribute to these disparities. These include lack of diversity in Science, Technology, Engineering, Mathematics, and Medicine (STEMM) fields; bias in healthcare delivery and access; underinsurance in patients from low-income backgrounds; and lack of training that advances those from historically underrepresented groups. The SCHEQ Foundation (STEMM & Cancer Health Equity) is a black-founded nonprofit organization that seeks to increase STEMM workforce diversity and improve outcomes for underrepresented, underserved, marginalized, and invisible populations across the cancer care continuum. Herein we describe the formation of SCHEQ Foundation and our core domains that we seek to make change. We seek to increase STEMM access and exposure, increase STEMM workforce diversity, and improve outcomes for underserved populations navigating the cancer care continuum. This will be accomplished in part by 1) engaging professional in STEMM from their lived background and experiences, 2) developing training, mentoring, and professional developing programs across the academic continuum to increase success of minoritized scholars; and 3) providing information and resources in culturally responsive manners to help underserved patient navigate cancer and advocate for the standard of care they deserve. We will change how we convey content, information, and resources to underserved populations by utilizing a combination of multimedia, infographics, webinars, info sheets, and community engagement. The goal is to increase survival for groups that historically have the greatest cancer disparities.

Citation Format: Eugene Manley. SCHEQ Foundation (STEMM & Cancer Health Equity): Increasing STEMM diversity and outcomes for patients navigating 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 1314.
Abstract 1339: Pre-clinical development of IAMA-004, a tri-specific antibody targeting innate immune receptors and tumor targets
Authors: Liu Liming , Han Zhen , Fan Yang , Pan Qi
Published: March 2024

Immune check-point inhibitor (ICI) such as anti-PD1 or anti-PDL1 antibody, which enhances the adaptive immunity via re-energizing exhausted T cells, has become the foundational therapy for many cancers. However, due to variety of other immunosuppressive tumor microenvironments, it is efficacious only in 15-30% of the patients. It is well known both the innate immunity and the adaptive immunity are required for full control of cancer growth. Thus, therapeutics harnessing the innate immunity will be complementary to ICI and improve the cancer response rate when used in combination. Currently therapeutics harnessing the innate immunity mainly involve bispecific antibodies with one arm targeting tumor antigen and one arm connecting to innate effector cells such as NK cells or macrophages, represented by CD30-CD16a bispecific and Her2-CD47 bispecific respectively. Considering that cancer patient’s immune status are heterogenous, varying from T cell exhaustion, macrophage disfunction (M2 bias), Treg cell presence to NK cell deficiencies, it would be advantageous to combine several innate immunity enhancers into one therapeutic molecule. Here we describe a long-acting tri-specific antibody termed IAMA-004 that targets Her2 as the primary target and CD47 and CD16a as effector cell engagers. CD47 is also a co-target on some tumor cells (CD47-Her2 double positive tumors). We have characterized this molecule both in vitro and in vivo. The in vitro experiments tested the molecule’s ability to kill Her2 high, Her2 low and Her2 negative tumor cell lines including BT-474, SK-OV-3, NCI-N87, OE19, HCC1954, MDA-MB-231, MDA-MB-468, in human PBMC ADCC assays with DS-8201 and Trastuzmab as the comparator molecules. In all of these ADCC experiments, IAMA-004 showed superior tumor cell killing activities to those mediated by DS-8201 or Trastuzmab. When combined with IAMA-005 (a Fc-IL2-IFN-α fusion cytokine), the tumor cell killing activities were dramatically increased. In addition, IAMA-004 was also tested in patient-derived Ex-Vivo fluids (one malignant pleural effusion and one malignant asities) and demonstrated tumor cell killing efficacies; and the killing activity could be attributed to the specific arms of the molecule. These results suggest that it is feasible to combine more than one innate engager in one molecule to carry out specific functions in the heterogeneous immune status for the treatment of advanced cancer patients.

Citation Format: Liming Liu, Zhen Han, Yang Fan, Qi Pan. Pre-clinical development of IAMA-004, a tri-specific antibody targeting innate immune receptors and tumor targets [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 1339.
Abstract 1354: Anti-CTLA-4 generates memory T-cells with greater expansion and functionality than anti-PD-1
Authors: Mok Stephen , Liu Huey , Anang Nana-Ama A.S. , Mancuso James J. , Çobanoğlu Didem Ağaç , Allison James P.
Published: March 2024

Distinct effects on T-cell differentiation arise from immune checkpoint inhibition targeting cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) and programmed death 1 (PD-1). However, the effect of these immunotherapies on the immunological memory response remains unclear. Our investigation into the effects of anti-CTLA-4 and anti-PD-1 on memory T-cell formation in mice revealed that anti-CTLA-4 generates a more effective memory antitumor response than anti-PD-1. Memory T-cells arising after anti-CTLA-4 treatment exhibit greater expansion, cytokine production, and antitumor activity than those from anti-PD-1. Notably, anti-CTLA-4 preserves more TCF-1+ T-cells during priming, while anti-PD-1 leads to more TOX+ T-cells. Experiments using conditional TCF-1- or TOX-knockout mice highlight TCF-1 is essential for memory response generated by anti-CTLA-4, whereas TOX deletion alone in T cells has no effect on response to anti-PD-1. Deepening our understanding of how checkpoint inhibition affects memory response is crucial for advancing our understanding of the enduring impacts of these immunotherapies on the immune system.

Citation Format: Stephen Mok, Huey Liu, Nana-Ama A.S. Anang, James J. Mancuso, Didem Ağaç Çobanoğlu, James P. Allison. Anti-CTLA-4 generates memory T-cells with greater expansion and functionality than anti-PD-1 [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 1354.
Abstract 1364: Characterization of CHS-1000, an Fc-modified anti-ILT4 monoclonal antibody for reprogramming suppressive myeloid cells in solid tumors
Authors: Rajasekaran Narendiran , Wang Xiaoguang , Klakamp Scott , Ravindranathan Sruthi , Tatarewicz Suzanna , Du Julia , Nguyen Brian , Widmann Kate , Hawkins Nessa , Chin Daniel J. , LaVallee Theresa , Kapoor Varun
Published: March 2024

Tumor resident immunosuppressive myeloid cells play a significant role in inducing resistance to cancer immunotherapy and are associated with poor prognosis in several solid tumors. The leukocyte immunoglobulin-like receptor B2 (LILRB2), also known as ILT4, is primarily expressed on myeloid cells and is a negative regulator of myeloid cell activation. ILT4 and its primary ligands (HLA-G, HLA-A) are widely expressed in various solid tumors and are associated with disease progression, tumor metastasis and poor clinical outcome. Abrogating inhibitory signals mediated through ILT4 and its ligands is a promising strategy to reprogram suppressive myeloid cells and enhance efficacy of immune checkpoint therapies. Here we report IND-enabling preclinical characterization of CHS-1000, a novel recombinant humanized Fc-modified IgG1 antibody with attenuated Fc-gamma receptor (FcγR) binding, that specifically targets immunoglobulin-like transcript 4 (ILT4). CHS-1000 selectively binds to human ILT4 with high affinity and potently blocks ILT4 interactions with its primary ligands (HLA-A, HLA-G). Functionally, CHS-1000 promotes re-polarization of suppressive myeloid cells to a pro-inflammatory phenotype and enhances activation of dendritic cells and T cells. Combinations of CHS-1000 with PD-1 inhibitors or other immunotherapy agents hold particular promise for enhanced anti-tumor activity. These preclinical characterization data support clinical development of CHS-1000 in combination with toripalimab (a commercial stage anti-PD-1 antibody) as an immunotherapy treatment for patients with solid tumors.

Citation Format: Narendiran Rajasekaran, Xiaoguang Wang, Scott Klakamp, Sruthi Ravindranathan, Suzanna Tatarewicz, Julia Du, Brian Nguyen, Kate Widmann, Nessa Hawkins, Daniel J. Chin, Theresa LaVallee, Varun Kapoor. Characterization of CHS-1000, an Fc-modified anti-ILT4 monoclonal antibody for reprogramming suppressive myeloid cells in solid tumors [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 1364.
Abstract 138: UCHL5 and PAX3-FOXO1 resolve replication stress in rhabdomyosarcoma
Authors: Sabnis Amit J. , Sahu Pushpendra K. , Pan Yue
Published: March 2024

PAX3-FOXO1 positive rhabdomyosarcoma (RMS) is an aggressive pediatric sarcoma with a terrible prognosis. The resultant fusion acts as a chimeric transcription factor, in part remodeling chromatin to inhibit terminal differentiation and maintain cancer cell proliferation. As this oncoprotein is not directly druggable, we have used genetic screens to identify dependencies downstream of PAX3-FOXO1 to improve our understanding of pathogenesis and develop new therapeutic targets. We performed pooled CRISPRi screening in an isogenic system of a patient-derived RMS cell line with or without knockdown of PAX3-FOXO1. Loss of the ubiquitin hydroxylase UCHL5 led to selective depletion of PAX3-FOXO1 positive Rh30 cells compared to PAX3-FOXO1 knockdown cells, suggesting that the fusion creates UCHL5 dependence. We validated this finding in competition growth assays in additional fusion positive (FP) and fusion negative (FN) RMS cell lines, and observed UCHL5 dependency only in the former. By contrast, UCHL5 was dispensable for cell growth and survival in FN RMS cell lines. These data were recapitulated in the large DepMap dataset, which also identified the top genetic co-dependencies of UCHL5 as INO80-complex (INO80c) members. We therefore investigated whether the fusion selective effects of UCHL5 were mediated by the INO80c, of which UCHL5 is an N-terminal member. We observed that knockdown of another N-terminal component of INO80c, NFRKB, phenocopies the fusion-selective effects of UCHL5 loss. Conversely, the catalytic ATPase INO80 was pan-essential, suggesting that only N-terminal subunits of INO80c regulate PAX3-FOXO1 specific functions. INO80c plays an essential role in mitigating DNA replication stress. Indeed, loss of UCHL5 led to DNA damage as measured by γH2AX and slowing of replication forks as measured by fiber assays in FP, but not FN RMS. Unexpectedly, acute degradation of PAX3-FOXO1 also slowed DNA replication fork speed, suggesting a genetic model in which UCHL5 and PAX3-FOXO1 cooperate to resolve replication stress in FP RMS. Finally, in vivo experiments using cell line xenografts confirms that loss of UCHL5 slows the growth of FP RMS, but not FN RMS. These findings suggest that loss of UCHL5 induces replication stress in a fusion-specific manner, and offers both an opportunity to study the mechanisms by which the PAX3-FOXO1 oncoprotein mitigates replication stress, and to develop UCHL5 inhibitors as precision therapies for FP RMS patients.

Citation Format: Amit J. Sabnis, Pushpendra K. Sahu, Yue Pan. UCHL5 and PAX3-FOXO1 resolve replication stress in rhabdomyosarcoma [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 138.
Abstract 1414: HBM9014, a first-in-class fully human anti-LIFR antibody with excellent preclinical efficacy and safety profile
Authors: Zeng Jue , Zhang Dongxu , Liu Louis , Zhang Lingbing
Published: March 2024

Background: Leukemia inhibitory factor (LIF) and cardiotrophin like cytokine factor 1 (CLCF1) are multi-functional cytokines from IL-6 family. They are reported to be highly expressed in a variety of human cancers such as pancreatic carcinoma, cholangiocarcinoma, NSCLC, GBM, head and neck cancer etc. Their expression is associated with poor prognosis and survival. LIFR is the key component of LIF/CLCF1 receptors. Antibody targeting LIFR has the advantage to block both LIF and CLCF1 pathways thus inhibiting tumor growth more effectively than products to block single cytokines.

Methods: Fully human Anti-LIFR antibodies were generated from H2L2 Harbour Mice® transgenic platform. In vitro function of the antibodies was tested for their abilities to block LIF/CLCF1 induced downstream signaling pathway and tumor spheroid proliferation. In vivo efficacy was tested in multiple tumor models as monotherapy and in combination with cisplatin. DRF toxicokinetics was evaluated on NHP with a 4-week repeat-dose toxicity study.

Results: HBM9014, a fully human anti-LIFR antibody generated by H2L2 Harbour Mice®, specifically binds LIFR thus blocks the interaction between LIF/CLCF1 and their respective receptors, inhibits downstream STAT3 phosphorylation, and reduces exogenous and endogenous LIF induced tumor spheroid proliferation in vitro. Furthermore, HBM9014 shows potent anti-tumor activity in multiple tumor models in vivo. In a GLP NHP 4-week repeat-dose toxicity study, HBM9014 is well tolerated up to 150 mg/kg with no discernable drug-related toxicity.

Conclusions: HBM9014 specifically binds to the key component of LIF and CLCF1 receptor, LIFR, thus blocks both LIF and CLCF1 signaling pathways. HBM9014 shows significant cell proliferation inhibition alone or in combo with small molecule inhibitors in 3D culture of multiple cancer cell lines. It also shows significant in vivo antitumor efficacy as monotherapy, and enhanced efficacy in combination with Cisplatin in multiple mouse models. Importantly, despite the benefits of double blocking, HMB9014 has an excellent safety profile. Taken together, HMB9014 is a promising drug candidate for advanced cancer with favorable efficacy and safety profiles that warrant clinical validation.

Citation Format: Jue Zeng, Dongxu Zhang, Louis Liu, Lingbing Zhang. HBM9014, a first-in-class fully human anti-LIFR antibody with excellent preclinical efficacy and safety profile [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 1414.
Abstract 1432: Development of transparent transgenic zebrafish model to study CNS microglia and NF-kB activity in glioblastoma
Authors: Rajpurohit Surendra K. , Nasanally Sai , Hotwani Simran , Gossman Taitum , Bhatt Keshu , Van Derhei Aiden , Pasi Rohan , Manhiani Devan-Anmol , Kimmerling Kirk , Lokeshwar Balakrishna L.
Published: March 2024

Microglia are resident macrophages of the central nervous system (CNS) and have crucial roles in normal brain homeostasis, function, and immune response. Microglia play key roles in the healthy brain, ranging from phagocytosis of dying neurons and CNS debris to modulating adaptability. The role of microglia has been well documented but their specific contributions to the development and progression of cancers, such as glioblastoma, have recently garnered wide attention. The zebrafish have proven novel tool for modeling innate immune responses. A macrophage-specific marker mpeg1 and its promoter are used in mpeg1-driven transgenes. mpeg1-driven transgenes mpeg1:mCherryFP are expressed in macrophage-lineage cells that do not express neutrophil-marking transgenes. These zebrafish transgenes contribute to the fields of inflammation, infection, and leukocyte biology. We have recently developed a novel transparent transgenic homozygous strain Tg(6xNF-kB:EGFP); Casper(roy−/−, nacre−/−) of zebrafish to the in-vivo study of NF-kB activity under normal and inflammation stage at the cellular, organ-specific and whole organism level. To perform an in-vivo study of microglial function, we are developing Casper;mpeg1:mCherryFP microglia specific strain and integrating with newly developed Tg(6xNF-kB:EGFP); Casper(roy−/−, nacre−/−) strain through crossbreeding of Casper and transgenic zebrafish. We have generated F1 generation of newly developed strains of Casper;mpeg1:mCherryFP and Tg(6xNF-kB:EGFP);Casper. The presence of fluorescence mCherry and GFP, in crossbred progeny was identified through the screening of larvae at 72 HPF stage. The expression of these fluorescent proteins determines the presence of the reporter genes. F1 generation confirmed heterozygous and allowed them to grow into adults to breed transparent transgenic strain to get F2 generation. This process will be continued to F2, F3, F4, and F5 generation until achieving homozygous progeny. This model allows for in-vivo examination of microglia functional morphology by using fluorescent and confocal microscopy. This novel transgenic model will be applied to identify novel anti-inflammatory drug inhibitors in NF-kB pathophysiology for glioblastoma and cardio-oncology experimental therapeutics.

Citation Format: Surendra K. Rajpurohit, Sai Nasanally, Simran Hotwani, Taitum Gossman, Keshu Bhatt, Aiden Van Derhei, Rohan Pasi, Devan-Anmol Manhiani, Kirk Kimmerling, Balakrishna L. Lokeshwar. Development of transparent transgenic zebrafish model to study CNS microglia and NF-kB activity in glioblastoma [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 1432.
Abstract 1436: Cancer associated fibroblast dependency in a model of bladder cancer
Authors: Valentine Henkel , Satyal Uttam , Bukavina Laura , Uddin Hassan , Beyamin Yousttena , Arya Rhea , Abbosh Philip
Published: March 2024

Introduction: The relationship between genotype and phenotype in targeted therapy has proven to be crucial for most kinase inhibitors. In bladder cancer, the PI3K/AKT/mTOR pathway is widely activated by mutation but untargeted. Our study focuses on developing an immunocompetent, CRISPR-edited bladder cancer mouse model that mirrors common PI3K/AKT/mTOR mutations. We aim to establish genotype-effective kinase inhibitor pairings using in vitro CRISPR-edited organoids and Cancer Associated Fibroblasts (CAFs) to create a pro-tumorigenic environment in vivo.

Methods and Results: Urothelial cells were isolated from the bladder of Rosa26 Lox-Stop-Lox-Cas9/GFP transgenic mice to develop organoids. These organoids were co-edited for Trp53 and Rb1 KO, and further editing of the Pten gene resulted in a nine-fold increase in sensitivity to the kinase inhibitor ipatasertib. Despite successful engraftment into mouse bladder, these edited cells regressed before tumor formation, and intradermal injection did not yield subcutaneous tumors. An alternate model featuring a 9p21 deletion with Trp53 KO also displayed similar regression post-engraftment into bladder or skin. However, co-injection with skin-cancer-derived CAFs subcutaneously yielded palpable subcutaneous tumors that grew for 12 weeks. Enhancing orthotopic model fidelity, we established a BBN bladder-tumor-derived CAF line, leading to the formation of tumors with squamous differentiation upon intramural co-injection with edited organoids. These models displayed immune-excluded architecture and a predominance of M2 over M1 macrophages. Following our accomplishments in establishing bladder tumors, we aimed to enhance our model's adaptability by introducing common PI3K/AKT/mTOR mutations, specifically TSC1/TSC2 KO and Pik3ca hotspots, into our 9p21/Trp53 KO model. While TSC1/TSC2 KO lines exhibited elevated P-S6K signaling, they displayed resistance to Torin 1, Voxtalisib, and Everolimus—kinase inhibitors targeting mTORC1/2. In parallel, we employed CRISPR-mediated HDR to successfully introduce a Pik3ca H1047R hotspot into our 9p21/Trp53 KO model, as evidenced by increased Phosphorylation of AKT compared to wild-type 9p21/Trp53 KO cell lines.

Conclusions: Our research establishes the crucial role of CAFs in bladder tumor formation. Our now established customizable model of bladder cancer is suitable for direct targeting by kinase inhibitors while providing a platform for understanding the genotype-phenotype relationship in bladder cancer.

Citation Format: Henkel Valentine, Uttam Satyal, Laura Bukavina, Hassan Uddin, Yousttena Beyamin, Rhea Arya, Philip Abbosh. Cancer associated fibroblast dependency in a model of bladder 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 1436.
Abstract 1465: Studying basal cell adaptation in the airways in response to chronic tobacco exposure
Authors: Peña David Osuna de la , Stoichkova Zornitsa , Przybilla Moritz Jakob , Percival Charlotte , Clarke Sarah E. , Gowers Kate H. , Campbell Peter J. , Janes Sam M.
Published: March 2024

Premalignant lesions in the airways are preceded by the accumulation of cells with a high mutational load, many of which carry driver mutations. Our previous work suggests this is a dynamic process: tobacco smoking increases the size of mutated clones, which contain shorter telomeres and higher Ki67, while smoking cessation appears to be associated with the expansion of clones with near-normal mutational burden and an overall reduction in Ki-67 expression. To understand how these long-term changes take place, we have performed ex vivo functional assays to quantify the competitive advantage imparted by smoke. We have then explored the transcriptomic landscape of these cultures to determine how differential smoke exposure modulates epithelial cell dynamics before and after smoking cessation.

Primary basal cells from bronchial explants were co-cultured following different regimes of exposure to cigarette smoke extract (CSE). We find that, upon acute exposure, pre-exposed cells have a ~20% higher chance of proliferating per week than smoke-naïve cells. To test this in a more physiological model, long-term epithelioid cultures were established with chronic exposure to low-dose CSE for months. As before, we observe that over time, pre-exposed cells outcompete the smoke-naïve, suggesting that smoke imparts a beneficial adaptation on re-exposed cells even at low doses. To elucidate this mechanism, we performed RNA sequencing of 48 different combinations of acute and chronic exposure to CSE. Genes were classified into distinct modules according to their regulation following exposure, re-exposure and cessation. Differential expression analysis and gene set enrichment analysis showed that metabolic pathways, including both nutrient and detoxification pathways, are predominantly enriched in these modules. In particular, following cessation, cells with the highest cumulative exposure to CSE displayed senescent-like features which are suppressed with chronic re-exposure. Further analysis and in vivo validation of these results will help elucidate how these changes affect the likelihood of premalignancy and may provide valuable targets for cancer prevention.

Citation Format: David Osuna de la Peña, Zornitsa Stoichkova, Moritz Jakob Przybilla, Charlotte Percival, Sarah E. Clarke, Kate H. Gowers, Peter J. Campbell, Sam M. Janes. Studying basal cell adaptation in the airways in response to chronic tobacco exposure [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 1465.
Abstract 1473: Exposure of arsenic promotes malignant stemness and induces bladder malignancy in preclinical models
Authors: Tyagi Bhawna , Vashishta Mohit , Tyagi Neha , Chandrasekaran Balaji , Shukla Vaibhav , Tyagi Ashish , Damodaran Chendil
Published: March 2024

Numerous epidemiological and scientific studies have consistently demonstrated that populations exposed to arsenic (As) concentrations ranging from >10 ppb to 2500 ppb in their drinking water face an elevated risk of urogenital cancers, with a particular emphasis on bladder cancer (BCa). Hence, the goal of the study is to investigate the underlying mechanisms that trigger malignant transformation in the bladder using physiologically relevant arsenic concentrations observed in BCa patients. In our results, we observed significantly elevated levels of arsenic in the urine of BCa patients, ranging from 20 to 50 mg/L, in stark contrast to the levels observed in healthy controls (>10 mg/L) and the recommended guidelines set by the Environmental Protection Agency (EPA) and Centers for Disease Control and Prevention (CDC). We used a median physiological concentration of arsenic (30 PPM) in drinking water-induced bladder hyperplasia in male and female mice over six months. Likewise, chronic exposure of healthy bladder epithelial cells (TRT-HU1) to similar arsenic concentrations (250nM:30 PPM) for 12 months resulted in malignant transformation, and arsenic-transformed cells (AsT) were capable of inducing tumors in xenografted and orthotopic models. Nonetheless, the molecular mechanism responsible for this malignancy remains to be elucidated. Our molecular analysis has pointed towards the activation of stem cells, notably ALDH1A1 (with a 50,000-fold increase), as a pivotal factor driving this transformation. Furthermore, our research has demonstrated that the activation of ALDH1A1 is contingent on the induction of Metal Response Element-Binding Transcription Factor-1 (MTF1). Consequently, silencing MTF1 inhibited the expression of ALDH1A1 and its associated malignant properties in As-transformed cells, strongly suggesting that MTF1 functions as the master regulator in As-induced bladder carcinogenesis. We have also observed similar molecular signatures (MTF1 and ALDH1A1 expression) in arsenic-exposed versus non-exposed mice and bladder cancer patients. In conclusion, our studies suggest that activation of stem cells is a critical step in arsenic-induced bladder carcinogenesis.

Citation Format: Bhawna Tyagi, Mohit Vashishta, Neha Tyagi, Balaji Chandrasekaran, Vaibhav Shukla, Ashish Tyagi, Chendil Damodaran. Exposure of arsenic promotes malignant stemness and induces bladder malignancy in preclinical models [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 1473.
Abstract 1548: Olfactomedin 4 deficiency promote de novo progression of prostate cancer in TRAMP mouse model
Authors: Li Hongzhen , Liu Wenli , Zhu Jianqiong , Chin Kyung , Rodgers Griffin P.
Published: March 2024

Prostate cancer is the most diagnosed solid tumor and the second leading cause of cancer-related death in American men. Olfactomedin (OLFM4) plays tumor suppressor roles and is associated with the initiation, growth, and clinical outcome of human prostate cancer. Olfm4-null mouse sporadically developed prostate epithelial neoplasia through up-regulated hedgehog signaling pathways. In this study, we have generated Olfm4+/+/GFP/TRAMP and Olfm4−/−/TRAMP mouse models to investigate Olfm4 gene biological functions underlying prostate cancer de novo progression. We have observed that Olfm4−/−/TRAMP mice developed more advanced prostate cancer at 47.6% (20/42) compared to Olfm4+/+/TRAMP at 11.9 % (5/42) and had poor survival compared to Olfm4+/+/TRAMP mice (P<0.01). Histopathological and immunofluorescent staining revealed that tumor tissues containing more heterogeneous cells such as Ar+/Ck8+ epithelial or Syn+ neuroendocrinal cells or Vim+/Ck8+ epithelium mesenchymal transition cells in the primary tumors obtained from Olfm4−/−/TRAMP mouse. We found more tumor angiogenesis and metastases into lymph nodes, liver, lung, kidney, and soft tissues in Olfm4−/−/TRAMP mice. The accumulations of the dendritic cell population in primary tumor and lung metastasis tumors were detected. The primary tumor was sensitive to androgen deprivation therapy by using castration with both Olfm4+/+/GFP/TRAMP and Olfm4−/−/TRAMP mice. However, castration-resisted EMT tumor cells or expressing neuroendocrine marker cells regrew after castration 7-8 months in both Olfm4+/+/GFP/TRAMP and Olfm4−/−/TRAMP mice. Furthermore, we tested JQ1, a BET bromodomain inhibitor, as an epigenetic targeted therapy reagent and found that JQ1 significantly inhibited tumor cell growth of Olfm4−/−/TRAMP mouse tumor in the organoids culture. Taken together, we have provided novel evidence that Olfm4 plays a tumor-suppressing role during prostate cancer de novo progression.

Citation Format: Hongzhen Li, Wenli Liu, Jianqiong Zhu, Kyung Chin, Griffin P. Rodgers. Olfactomedin 4 deficiency promote de novo progression of prostate cancer in TRAMP mouse model [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 1548.
Abstract 1554: LIM Domain 7 (LMO7) splice variant influences prostate cancer biology
Authors: Abo Muthana Al , George Daniel J. , Wang Zefeng , Patierno Steven R. , Freedman Jennifer A. , Jiang Alice
Published: March 2024

LIM Domain 7 (LMO7) has been shown to participate in cell-cell junction formation through interaction with afadin and α-actinin. It has also been reported to collaborate with Rho kinase to regulate actin filament formation and to activate the Rho-myocardin-related transcription factor serum response factor pathway. Additionally, LMO7 itself may function as a transcription factor, regulating the expression of genes involved in Epithelial Mesenchymal Transition (EMT). Alternative splicing of LMO7 has the potential to modulate its function. Previously, we reported LMO7 as a target that is alternatively spliced in prostate cancer (PCa) between self-reported Black and White patients. In addition, by analyzing expression of LMO7 splice variants in The Cancer Genome Atlas (TCGA) data, we previously reported increased expression of a LMO7 splice variant skipping exon 12 in breast, lung and liver cancer compared with paired normal tissues. In the present study, we found that LMO7 is downregulated in metastatic PCa specimens within the Stand Up To Cancer cohort. To investigate the functional impact of expression of the LMO7 splice variant skipping exon 12 in PCa, we utilized CRISPR-Cas9 methodology to delete LMO7 exon 12 in PCa cells and assessed resulting alterations in PCa cell biology. The LMO7 exon 12 deleted PC3 PCa cells exhibited enhanced migratory potential compared with parental PCa cells. An Immunofluorescence (IF) assay showed weak staining of cell-cell junctions in LMO7 exon 12 deleted PCa cells compared with parental PCa cells. Moreover, IF staining showed retardation in f-actin in LMO7 exon 12 deleted PCa cells compared with parental PCa cells. As E-Cadherin is often inhibited during EMT, we assessed the level of E-Cadherin by western blot and real-time polymerase chain reaction (PCR). These analyses revealed that E-Cadherin protein as well as messenger RNA is suppressed in LMO7 exon 12 deleted PCa cells compared with parental PCa cells. These findings suggest a role for skipping of LMO7 exon 12 in disrupting PCa cell-cell junctions, down-regulating E-Cadherin in PCa and enhancing PCa cell migratory potential, consistent with EMT.

Citation Format: Muthana Al Abo, Daniel J. George, Zefeng Wang, Steven R. Patierno, Jennifer A. Freedman, Alice Jiang. LIM Domain 7 (LMO7) splice variant influences prostate cancer biology [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 1554.
Abstract 1566: Cancer metastasis and stemness: The metastasis-inducer MACC1 regulates LGR5 to promote cancer stem cell-like properties in colorectal cancer
Authors: Erdem Müge , Lee Kyung Hwan , Hardt Markus , Regan Joseph , Kobelt Dennis , Walther Wolfgang , Regenbrecht Christian , Stein Ulrike S.
Published: March 2024

Colorectal cancer (CRC) is one of the leading causes of cancer-related deaths worldwide. The high mortality is directly associated with metastatic disease which is thought to be initiated by colon cancer stem cells according to cancer stem cell (CSC) model. Consequently, early identification of those patients who are at high risk for metastasis is crucial for improved treatment and patient outcome. Metastasis-associated in colon cancer 1 (MACC1) is a novel prognostic biomarker for tumor progression and metastasis formation independent of tumor stage. We previously showed an involvement of MACC1 in cancer stemness in the mouse intestine of our transgenic mouse models. However, the expression of MACC1 in human CSCs and possible implications remain elusive. Here, we explored the molecular mechanisms by which MACC1 regulates stemness and CSC-associated invasive phenotype based on patient-derived 3D cell culture models (PD3D), patient-derived xenografts (PDX) and human CRC cell lines. We showed that CD44-enriched CSCs from PD3D models express significantly higher levels of MACC1 and display higher tumorigenicity in immunocompromised mice. Similarly, RNA sequencing performed on PD3D and PDX models demonstrated significantly increased MACC1 expression in ALDH1(+) CSCs, highlighting its involvement in cancer stemness. We further showed the correlation of MACC1 with CSC markers CD44, NANOG and LGR5 in PD3D models as well as established cell lines. Additionally, MACC1 increased stem cell gene expression, clonogenicity and sphere formation. Strikingly, we showed that MACC1 binds as a transcription factor to the LGR5 gene promoter uncovering the long-known CSC marker LGR5 as novel essential signaling mediator used by MACC1 to induce CSC-like properties in human CRC patients. Our in vitro findings were further substantiated by significant positive correlation of MACC1 and LGR5 in CRC cell lines as well as CRC patient tumors. Taken together, this study indicates that the metastasis-inducer MACC1 act as a cancer stem cell-associated marker. Interventional approaches targeting MACC1 would potentially improve further targeted therapies for colorectal cancer patients to eradicate CSCs, prevent cancer recurrence and distant metastasis formation.

Citation Format: Müge Erdem, Kyung Hwan Lee, Markus Hardt, Joseph Regan, Dennis Kobelt, Wolfgang Walther, Christian Regenbrecht, Ulrike S. Stein. Cancer metastasis and stemness: The metastasis-inducer MACC1 regulates LGR5 to promote cancer stem cell-like properties in colorectal 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 1566.