bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract PR005: Targeting EP300 and CBP for therapeutic benefit in pediatric solid tumors
Authors: Durbin Adam D. , Shendy Noha , Mercier Audrey , Zhang Yang , Bikowitz Melissa J. , Sigua Logan H. , Robinson Sarah , Wang Tingjian , Jonchere Barbara , Look A. Thomas , Zimmerman Mark W. , Roussel Martine , Abraham Brian J. , Schonbrunn Ernst , Stegmaier Kimberly , Qi Jun
Published: December 2022

Gene expression is regulated by promoters and enhancers marked by histone H3 lysine 27 acetylation (H3K27ac), which is established by the paralogous, multidomain-containing histone acetyltransferases (HAT) EP300 and CBP. These enzymes display overlapping regulatory roles in untransformed cells, but less characterized roles in cancer cells. We demonstrate that the majority of high-risk pediatric neuroblastoma (NB) depends on EP300, whereas CBP has a limited role. To disrupt EP300, we developed a proteolysis-targeting chimera (PROTAC) compound termed “JQAD1” that selectively targets EP300 for degradation. JQAD1 treatment causes loss of H3K27ac at the enhancers that regulate NB master transcription factors, and drives rapid NB apoptosis, with limited toxicity to untransformed cells where CBP may compensate. In parallel, we demonstrate that EP300 and CBP have subdomain-specific functions, with enriched activity for individual domains in distinct tumor lineages. These data provide a foundation for interrogation of these histone acetyltransferases in distinct tumor types and new strategies for therapeutic disruption of subdomain and scaffolding activities of these multidomain proteins in cancer.

Citation Format: Adam D. Durbin, Noha Shendy, Audrey Mercier, Yang Zhang, Melissa J. Bikowitz, Logan H. Sigua, Sarah Robinson, Tingjian Wang, Barbara Jonchere, A. Thomas Look, Mark W. Zimmerman, Martine Roussel, Brian J. Abraham, Ernst Schonbrunn, Kimberly Stegmaier, Jun Qi. Targeting EP300 and CBP for therapeutic benefit in pediatric solid tumors. [abstract]. In: Proceedings of the AACR Special Conference: Cancer Epigenomics; 2022 Oct 6-8; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2022;82(23 Suppl_2):Abstract nr PR005.
Abstract A014: Identifying and overcoming resistance to RAS targeted therapy in colorectal cancer
Authors: Parsons Marie J. , Zafra Maria , Goswami Sukanya , Fernandez Maria Teresa Calvo , Wilkinson John E. , Dow Lukas E.
Published: December 2022

KRAS is the most frequently mutated oncogene in human cancer and a potent oncogenic driver. In colorectal cancer (CRC), pathogenic KRAS mutations are observed in up to 50% of cases, yet there is little understanding of how distinct mutations dictate tumor biology including initiation, progression and therapy response. The recent approval of targeted KRASG12C inhibitors brings promise to transform the therapeutic landscape for many patients who lack effective, personalized treatment. Clinically acquired resistance is already observed, highlighting the need to identify mechanisms of resistance and approaches to overcome them. The aim of this study is to understand how distinct Kras alterations drive tumorigenesis and identify reasons underlying the failure of KRAS inhibitors in CRC. To explore the clinical significance of distinct KRAS mutations in tumor development and therapy response, we used Cre- conditional (Lox- stop- Lox) Kras mutant mouse strains crossed to the colon specific promoter Fabp1-Cre and Apc(flox/flox) mice. In the absence of Apc, we demonstrated that distinct Kras mutations G12C and G13D drive unique phenotypes in the colon which demonstrates that subtle mutational changes can dramatically alter the physiological response in vivo, and make distinct KRAS alleles an attractive target for targeted therapeutics. In tumor bearing models of CRC, we observed that KrasG12D drives large numbers of smaller tumors, while KrasG13D drives smaller numbers of larger tumors. To investigate the differences observed in tumorigenesis, we are studying the potential role of Kras to prime the colon for APC loss and the consequences of Kras alterations on the stem cell dynamics in driving tumorigenesis. Further, we employed our unique LSL-Kras mutant mouse strains and colon derived organoids to assess the therapeutic response of G12C and pan Kras inhibitors alone or in combination with upstream target drugs including inhibitors targeting EGFR signaling (EGFR, SHP2, SOS1 and MEK) and the Wnt signaling pathway. Together, this work has identified that distinct mutations in KRAS creates colon cancer specific vulnerabilities which dictates tumor biology. The unique in vivo and ex vivo organoid models provide an ideal platform to identify the underlying cause of failure of G12C inhibitors and developing effective approaches to target RAS in CRC.

Citation Format: Marie J. Parsons, Maria Zafra, Sukanya Goswami, Maria Teresa Calvo Fernandez, John E. Wilkinson, Lukas E. Dow. Identifying and overcoming resistance to RAS targeted therapy in colorectal cancer [abstract]. In: Proceedings of the AACR Special Conference on Colorectal Cancer; 2022 Oct 1-4; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;82(23 Suppl_1):Abstract nr A014.
Abstract A034: Cancer and immunology: The homeostasis dance
Authors: Ekine-Afolabi Bene Akromaa
Published: December 2022

The human body is made up of not only tissues and blood, but also present are microbiota on surfaces of the skin, mucosal membrane of the intestine, urogenital, lungs, and mouth, to form ecological niches. Some of these niches have been well studied, while some are yet to be elucidated due to the unknown presence of these microbiota in such system. T cell response by immune system is one step of the cancer-immunity cycle, that maintains the prevention of autoimmunity. Cancer cells have T cell inhibitory signals including PD-L1 which have been identified for the development of new immunotherapy that are specifically responsible for hindering immune effector inhibition, thereby reinvigorating, and enhancing pre-existing anticancer immune response. Previous activity in immune therapies have always considered suppressive factors in the tumor microenvironment without consideration to other factors such as the genetic basis of the immune system. Attention to the immune system has always been on the response to the pathogens, or the threatening foreign target, but not on the genes responsible for regulating the immune system. The immune system is a concert of interactions between existing microbes and host. One of the major tools of cellular interaction is epigenetics. Epigenetic information regulates differentiation and development, thus can impact on pathological condition. Therefore, epigenetics interaction between microbes and host is vital in cancer development and treatment. Diagnostic and prognosis will be impacted by the understanding of the interactions between microbes and host, thereby informing on precision medicine.

Citation Format: Bene Akromaa Ekine-Afolabi. Cancer and immunology: The homeostasis dance [abstract]. In: Proceedings of the AACR Special Conference on Colorectal Cancer; 2022 Oct 1-4; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;82(23 Suppl_1):Abstract nr A034.
Abstract B003: Cross-reacting antigens between the intestinal microbiome and tumor neoantigen are associated with immune checkpoint inhibitor response in a gastrointestinal tumor cohort
Authors: Poudel Ravin , Dai Die , Li Fang , Yin Xiaochen , Kou Yan , Gomes Antonio L. C.
Published: December 2022

Background: Immune checkpoint inhibitor (ICI) is a revolutionary therapy that promotes the activation of effector T cells against tumors. Despite the potential for long-term benefit, a large proportion of patients does not respond to ICI therapy. One reason for this heterogenic response is due to the high variability in the intestinal microbiota among different patients. The abundance of certain bacteria has been associated with different responses and fecal meta transplant from responding donors was shown to rescue a beneficial phenotype in refractory patients. The presence of cross-reactive antigens (CRAgs) between the gut microbiome and tumor neoantigens has been proposed as a potential mechanism that could link the intestinal microbiota with ICI response. However, empirical evidence of CRAg’s role in ICI therapy is limited to a few studies. Methods: We investigated ICI response in a cohort of 22 stage IV gastrointestinal (GI) tumor patients in which whole-exome and gut metagenomic sequencing data were available. This cohort included patients with gastric cancer (n=11), esophageal carcinoma (n=6), and colon cancer (n=5). For each patient, we characterized the tumor neoantigen load based on epitopes that show high affinity and presentability in HLA molecules. Neoantigens were matched to the intestinal microbiome to identify corresponding CRAgs. CRAg quality, which ranks CRAgs according to their immunogenic potential, was defined based on matches in immunogenic protein or epitope databases. Results: We found the total number of CRAgs varied per tumor type, with a higher abundance in colon cancer (median µ=619) followed by esophageal (µ =287.5) and gastric (µ = 243). Interestingly, colon cancer patients showed a better progression-free survival (PFS) curve followed by esophageal and gastric cancer patients, respectively. We also observed that CRAg load was better associated with longer PFS (p=.03) than neoantigen load (p=0.07). Moreover, CRAg’s quality showed to be an important predictor of PFS. Patients with a high proportion of CRAgs matching at least one of the quality databases showed better PFS (p=0.02). Further, a taxa-specific analysis indicated that commensal CRAgs load was a better predictor of PFS (p<0.008) than relative abundance. Altogether, our work supports a mechanistic link between the microbiome, neoantigens, and immuno-therapy.

Citation Format: Ravin Poudel, Die Dai, Fang Li, Xiaochen Yin, Yan Kou, Antonio L. C. Gomes. Cross-reacting antigens between the intestinal microbiome and tumor neoantigen are associated with immune checkpoint inhibitor response in a gastrointestinal tumor cohort [abstract]. In: Proceedings of the AACR Special Conference on Colorectal Cancer; 2022 Oct 1-4; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;82(23 Suppl_1):Abstract nr B003.
Abstract B032: Native sgRNA capture and sequencing (NSC-seq) reveals tumor formation in ApcMin/+ mice and in human colorectal cancer is an oligoclonal process
Authors: Islam Mirazul , Yang Yilin , Shah Vishal M. , Simmons Alan J. , Xu Yanwen , Coffey Robert J. , Lau Ken S.
Published: December 2022

CRISPR-Cas9-based technologies have been developed to address a wide range of biological questions, including molecular recording of embryonic development, genetic screening, and tracking tumor evolution at the single-cell resolution. We developed a single-cell lineage tracking platform called native sgRNA Capture and sequencing (NSC-seq) to leverage CRISPR technology for clonal analysis of intestinal tumors in the ApcMin/+ mouse model. We combined CRISPR-induced mutations with spontaneously accumulating mitochondrial variants to enable highly efficient lineage-tracking of single cells. We generated lineage-tracked ApcMin/+ tumors and found that there are more than two independently arising clones within a tumor. We also found oligoclonal tumors within human colorectal specimens using this technology, supporting the oligoclonality of intestinal tumor development across species. Finally, we demonstrated the application of this technology in understanding the rate of loss-of-heterozygosity events in Apc as related to the cellular turnover rates of different intestinal regions. Overall, NSC-seq enables in vivo molecular recording of spontaneous intestinal tumor development at single-cell resolution.

Citation Format: Mirazul Islam, Yilin Yang, Vishal M. Shah, Alan J. Simmons, Yanwen Xu, Robert J. Coffey, Ken S. Lau. Native sgRNA capture and sequencing (NSC-seq) reveals tumor formation in ApcMin/+ mice and in human colorectal cancer is an oligoclonal process [abstract]. In: Proceedings of the AACR Special Conference on Colorectal Cancer; 2022 Oct 1-4; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;82(23 Suppl_1):Abstract nr B032.
Abstract IA018: Microbiome and intestinal barriers in colon cancer
Authors: Sun Jun , Zhang Jilei , Xia Yinglin , Zhang Yong-Guo
Published: December 2022

Multiple factors (e.g., risk genes, environmental triggers, immunity, and microbiome) contribute to the pathogenesis of colorectal cancer (CRC). The intestinal dysbiosis and disruption of barrier function are common manifestations of CRC. Vitamin D/Vitamin D receptor (VDR) is known to protects against dysbiosis and tumorigenesis. Considering the multiple functional roles of VDR in the development of cancer, it is important to dissect the mechanisms by which host factors contribute to the intestinal barrier function in protecting cells from tumorigenesis. Here, we reported that in human tissue, reduced VDR in the colon of CRC patients is associated with the reduced intestinal barrier and enhanced bacterial translocation to tumors. VDR deficiency in the intestinal epithelial cells led to the reduced thickness of mucus layer and enhanced penetration of microbiota into the epithelial cells. VDR was a transcriptional factor regulating the genes of some tight junction (TJ) Claudins. The positively correlated status of VDR and Claudins (e.g., Claudin- 5 and10) contribute to the altered intestinal permeability in tumorigenesis. In the intestinal epithelial VDR conditionally knockout VDR∆IEC mice and myeloid VDR deletion mice, dysbiosis and inflammatory cytokines in the VDR deficient context further altered TJ proteins and manipulated the barrier functions in the development of colon cancer. Furthermore, we found that mice with colonic adenomatous polyposis coli (APC) mutation reduced thickness of mucus layer and enhanced penetration of microbiota into the epithelial cells, thus enhancing tumorigenesis in the colon. Studies on host factor regulation of microbiome and barriers could be potentially applied to risk assessment, early detection, and prevention of colon cancer. These studies will provide an additional avenue to restore microbiome and barrier functions for risk assessment and prevention of CRC and other cancers.

Citation Format: Jun Sun, Jilei Zhang, Yinglin Xia, Yong-Guo Zhang. Microbiome and intestinal barriers in colon cancer [abstract]. In: Proceedings of the AACR Special Conference on Colorectal Cancer; 2022 Oct 1-4; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;82(23 Suppl_1):Abstract nr IA018.
Abstract IA025: Stem and metaplastic cell states and their influences on the tumor immune microenvironment in colonic pre-cancerous lesions
Authors: Lau Ken S.
Published: December 2022

The immune microenvironment is critical to colorectal cancer (CRC) pathogenesis, regarding both prognosis and therapeutic responses. While much work has been done to characterize tumor microenvironments, less is known about those environments are established following transformation of epithelial cells. We hypothesize that mapping routes towards CRCs, from normal human colonic tissues and precursor polyps, will uncover additional mechanisms that define the CRC cellular landscape and identify targets with diagnostic or therapeutic utility. Here, we present a comparative atlas of the two most common colorectal precursor lesions, conventional adenomas and serrated polyps, and their resulting CRC counterparts using single-cell transcriptomics and multiplex imaging across two cohorts of human participants. Integrative analysis reveals adenomas arise from dysregulated WNT-driven expansion of stem cells, while serrated polyps are depleted of stem cell populations and are derived from differentiated cells transitioning through a pyloric metaplasia program. Mucosal damage associated with metaplasia is coupled to a cytotoxic immune microenvironment that precedes hypermutation. Mouse and organoid experiments validate the importance of the differentiation states of colonic tumor cells in driving cytotoxic immunity partly through increased expression of antigen-presentation machinery. These data provide constitute a rich resource into malignant progression of colorectal polyps and their microenvironments, serving as a framework for precision surveillance and prevention of CRC.

Citation Format: Ken S. Lau. Stem and metaplastic cell states and their influences on the tumor immune microenvironment in colonic pre-cancerous lesions [abstract]. In: Proceedings of the AACR Special Conference on Colorectal Cancer; 2022 Oct 1-4; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;82(23 Suppl_1):Abstract nr IA025.
Abstract IA027: Recent advances in the study of extracellular vesicles and nanoparticles in colorectal cancer
Authors: Coffey Robert J.
Published: December 2022

Extracellular vesicles (EVs) and nanoparticles (NPs) are increasingly recognized as important mediators of intercellular communication with implications for the diagnosis, monitoring and possibly treatment of colorectal cancer. We recently reviewed this topic in Gastroenterology. EVs are lipid bilayer-enclosed vesicles that can be classified as small (s) some of which are tetraspanin-containing, endosomally-derived exosomes (5-120 nm) and large that include plasma membrane-budding microvescles (150-800 nm). Nanoparticles are more recently discovered smaller amembranous nanoparticles that include exomeres (30-50 nm) and supermeres (25-35 nm). We have performed a comprehensive proteomic and RNA analysis of sEVs, exomeres and supermeres. Highlights from this analysis will be presented with a focus on their clinical relevance and the merits of parsing versus pooling biofluid fractions.

Citation Format: Robert J. Coffey. Recent advances in the study of extracellular vesicles and nanoparticles in colorectal cancer [abstract]. In: Proceedings of the AACR Special Conference on Colorectal Cancer; 2022 Oct 1-4; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;82(23 Suppl_1):Abstract nr IA027.
Abstract PR001: The use of risk prediction for patients with abnormal fecal testing (FIT) to prioritize patient navigation
Authors: Petrik Amanda F.
Published: December 2022

Context: “Precision delivery involves using an individual’s electronic health data to predict risk and personalize care to substantially improve value” (Parikh). Rates of completion of the follow-up colonoscopy following an abnormal FIT are low in community clinics. Tools like predictive analytics can be used for population management and more precise delivery of care to those who need it the most. Objective: We describe 3 studies use of predictive analytics after abnormal FIT testing to prioritize patient navigation to complete subsequent colonoscopies. Study design and analysis: The Strategies and Opportunities to Prevent Colorectal Cancer (STOP CRC) trial developed an initial risk prediction model identifying patients unlikely to complete a follow-up colonoscopy on their own. Kaiser Permanente Northwest (KPNW) then redeveloped the model for use with Navigation. The Predicting and Addressing Colonoscopy Non-adherence in Community Settings (PRECISE) study then redeveloped the model for use with Navigation in a large Federally Qualified Health Center (FQHC). Results: The STOP CRC model included 8 variables, 1596 patients had an abnormal FIT result, 34.8% had a recorded colonoscopy within 6 months. The model shows an adequate separation of patients across risk levels for non-adherence to follow-up colonoscopy (bootstrap-corrected C-statistic > 0.63). The KPNW model included 14 variables, and the performance of the model was adequate with a bootstrap corrected C-statistic of 0.65. The PRECISE model included 12 variables and a bootstrap corrected C-statistic of 0.58. Here we will describe each of the models and the differences in the application of the models. Conclusions: Risk prediction can be used to identify patients unlikely to complete colonoscopies, and as a population management tool to tailor interventions to specific populations. Tailoring interventions can save money, and increase patient satisfaction.

Citation Format: Amanda F. Petrik. The use of risk prediction for patients with abnormal fecal testing (FIT) to prioritize patient navigation [abstract]. In: Proceedings of the AACR Special Conference on Colorectal Cancer; 2022 Oct 1-4; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;82(23 Suppl_1):Abstract nr PR001.
Abstract A003: Identification of a potential chemoprevention agent for BRCA1- mutated cancers
Authors: Shanmugam Sujeeth , Hosch Eboni , Konar Dipasri , Ford James M. , Alli Elizabeth
Published: December 2022

BRCA1 mutation carriers are predisposed to developing aggressive breast, ovarian, and other cancers at a relatively early age. Unfortunately, these cancers lack an effective chemoprevention option. We have found that cells containing mutant BRCA1 harbor defective repair of oxidative DNA damage, an early trigger to tumorigenesis. Therefore, we sought to target this defect and discover a novel chemoprevention agent for these cancers. High-throughput screening of a chemical library identified a novel class of small-molecules that enhance repair of oxidative DNA damage, which we termed DNA repair-activating agents. One of these agents, benserazide, increased base-excision repair of oxidative DNA damage utilizing a cell-based DNA repair assay, decreased levels of 8-oxo-g (i.e. most common oxidized lesion in human genome) using flow cytometric analysis, and decreased levels of oxidative DNA damage as determined by alkaline comet assay modified for detection of oxidized DNA lesions. These results were observed in mutant but not wild-type BRCA1 breast cancer cells. Benserazide, but not tamoxifen (current FDA-approved breast cancer chemoprevention agent), also decreased in vitro tumorigenesis of mutant BRCA1 or BRCA1-depleted breast cancer cells according to the soft-agar colony formation assay, and delayed tumor formation in vivo using a xenograft mouse model. Lastly, benserazide led to elevated levels of 8-oxo-dG (a by-product of base-excision repair) in conditioned media of mutant BRCA1 breast cancer cells, and concurrently increased serum levels of 8oxodG and decreased tumor burden in mice. Taken together, benserazide is a DNA-repair activating agent that enhances repair of oxidative DNA damage and decreases tumorigenesis in vitro and in vivo, and thus may serve as a potential chemoprevention agent for BRCA1-mutated cancers.

Further, circulating levels of 8oxoG/8oxodG may function as a predictive biomarker for the efficacy of benserazide, which would prove useful in the clinical evaluation of benserazide for cancer chemoprevention.

Citation Format: Sujeeth Shanmugam, Eboni Hosch, Dipasri Konar, James M. Ford, Elizabeth Alli. Identification of a potential chemoprevention agent for BRCA1- mutated cancers [abstract]. In: Proceedings of the Second Biennial NCI Meeting: Translational Advances in Cancer Prevention Agent Development (TACPAD); 2022 Sep 7-9. Philadelphia (PA): AACR; Can Prev Res 2022;15(12 Suppl_2): Abstract nr A003.
Abstract A011: Effect of IL-23 knockdown on obesity driven CRC in high-fat diet fed Apcmin/+ mice
Authors: Madka Venkateshwar , Chiliveru Srikanth , Zhang Yuting , Stratton Nicole , Bao Anh , Kumar Nandini , Rao Chinthalapally V.
Published: December 2022

Colorectal cancer (CRC) is the 3rd common cancer in the United States with an estimated 151,030 new cases and 52,580 deaths in 2022. Several studies suggest that western-style diet-induced obesity contributes to CRC promotion via systemic inflammatory mediators. Recently, we have reported significant elevation of circulating IL-23 levels in obese human subjects (BMI>30). Also, TCGA gene expression data indicated IL-23A over expression in colonic tumors (CT) that correlated with tumor stage, patient obesity, nodal metastasis, and poor survival rates. Importantly, we have shown that IL-23 knockdown in Apc min/+ mice led to significant suppression of the intestinal tumors strongly supporting our hypothesis that pro-inflammatory cytokine IL-23 could be a possible link between obesity and CRC. Here we explored the contribution of IL-23 in obesity induce CRC in vivo using high-fat diet (HFD) fed Apc min/+ mouse model. For this study, Apc min/+ and IL-23 KO (IL-23A p19-/- mice; MMMRC) were cross-bred to generate Apc min/+ mice with IL-23 normal (IL-23+/+) or KO (IL-23-/-) genotypes. Six-week old Apc min/+ mice (N≥15/gender) were grouped by IL-23 genotypes and fed HFD (60 Kcal) until termination. Mice were euthanized at 20 weeks age, intestines were harvested and evaluated for tumors incidence and multiplicity. In Apc min/+ mice, IL-23 deletion had significant suppressive effect on HFD driven CT and small intestinal tumors (SIT). IL-23-/-

Apc min/+ male mice had 50% less CT (0.81+0.25 vs 1.61±0.27; p<0.05) while female mice had 40% less CT (0.40±0.16 vs 0.94±0.21; p=0.05) compared to the IL-23+/+ Apc min/+ control mice. Similarly, CT incidence was also suppressed by 36% and 48% in male and female IL-23-/- Apc min/+ mice respectively compared to control mice. SIT multiplicity was also inhibited by 48% in male (15.00±1.13 vs 28.54±1.11; p<0.0001) and by 41% in female (14.83±1.35 vs 25.18±1.71; p<0.0005) IL23-/- Apc min/+ compared to control mice. Tumor data was complemented by significant reduction in markers of proliferation, immune evasion, circulating proinflammatory cytokine and chemokines (IL-1, IL- 17, IL-23, IL-10, CCL-3, CCL-2, CCL-5, IFNγ, TNFα, etc.) in IL-23 KO compared to control mice. Our data clearly indicates that IL-23 is a promising target for CRC prevention in high-risk obese individuals and warrants further investigation. (Supported in part by P30CA225520 and Kerley-Cade Endowed Chair).

Citation Format: Venkateshwar Madka, Srikanth Chiliveru, Yuting Zhang, Nicole Stratton, Anh Bao, Nandini Kumar, Chinthalapally V. Rao. Effect of IL-23 knockdown on obesity driven CRC in high-fat diet fed Apcmin/+ mice [abstract]. In: Proceedings of the Second Biennial NCI Meeting: Translational Advances in Cancer Prevention Agent Development (TACPAD); 2022 Sep 7-9. Philadelphia (PA): AACR; Can Prev Res 2022;15(12 Suppl_2): Abstract nr A011.
Abstract A11: Tumor-induced immune disorder: the root cause of cancer cachexia and therapeutic target
Authors: Zhang Lingbing
Published: December 2022

It’s well known that tumor growth induces immune system dysfunction, and cancer immunology has been focused on studying anti-tumor immunity, however the impact of the immune dysfunction on health of the host has not been studied. In this study, we investigate the immune system change and its impact on the host under tumor development from tumor establishment to overall health deterioration and death. We identified significant and multifaceted abnormalities of the immune system which triggers cancer cachexia onset and development. Thus, we propose a new term tumor-induced immune disorder (TID) for these abnormalities. We defined the TID by two parameters: multiple inflammatory cytokines expression; loss of balance and normal behavior of immune cells. Furthermore, we took efforts to explore potential treatments to correct the TID. We have identified a novel immunomodulator, i.e., (R)-ketorolac which can suppress expression of multiple inflammatory cytokines, and consistently, it can fully remove symptoms of cancer cachexia. Thus, we established the causative relationship between TID and cachexia. Interestingly cachectic mice treated by (R)-ketorolac are not only cured of cachexia thus avoid imminent death but also tolerate and respond to full dose anti-tumor chemotherapies. These findings have important clinical implications given majority of late-stage cancer patients with cachexia cannot tolerate chemotherapies and other anti-tumor treatments, thus get stuck in the treatment dilemma. Thus, this study points out a new direction to develop cancer immunotherapies by targeting and correcting the TID, which can not only prevent imminent death by curing cancer cachexia, but also provide opportunities for patients to receive and respond to anti-tumor treatments.

Citation Format: Lingbing Zhang. Tumor-induced immune disorder: the root cause of cancer cachexia and therapeutic target [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy; 2022 Oct 21-24; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(12 Suppl):Abstract nr A11.
Abstract A13: Siglec-7/9-sialic acid interactions inhibit T cell immune response in prostate cancer
Authors: Wen Ru M , Stark Jessica C. Stark C , García-Marqués Fernando , Zhao Hongjuan , Nolley Rosie , Bertozzi Carolyn R , Pitteri Sharon J , Brooks James D.
Published: December 2022

Immunotherapy has rapidly expanded in the care of patients with cancer with the discovery of immune checkpoints like PD-1 and CTLA-4. However, current immune checkpoint inhibitors are largely ineffective for prostate cancer. Recent studies suggest an alternative immune evasion pathway through the interactions between Sialic acid-binding immunoglobulin-type lectin proteins (Siglec) and their ligands, sialylated glycoprotein. Siglec-7/9-sialic acid interactions are reported to inhibit the immune cell response in several cancer types including leukemia, melanoma, and non-small cell lung cancer. Here, we demonstrated that Siglec-7/9 ligands were expressed in both prostate cancer tumor tissues and cell lines. We promoted T cell-mediated cytotoxicity of cancer cells by disrupting the interactions between Siglec-7/9 and their ligands. We discovered that FXYD5 and CD59 are potential Siglec-7 and Siglec-9 ligands, respectively, with CRISPRi screen. We then found that FXYD5 and CD59 knockout cells had reduced Siglec-7/9 binding capacity and enhanced T cells mediated killing effects on prostate cancer cells. These results provide a rationale for novel immune checkpoints and potential approaches for targeting Siglec-7/FXYD5 and Siglec-9/CD59 immune checkpoints for prostate cancer.

Citation Format: Ru M Wen, Jessica C. Stark C Stark, Fernando García-Marqués, Hongjuan Zhao, Rosie Nolley, Carolyn R Bertozzi, Sharon J Pitteri, James D. Brooks. Siglec-7/9-sialic acid interactions inhibit T cell immune response in prostate cancer [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy; 2022 Oct 21-24; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(12 Suppl):Abstract nr A13.
Abstract A43: Analysis of the effects of therapeutic vaccination with tumor vascular-primed dendritic cells in a mouse model of colon cancer
Authors: Wooster Amanda L , Anderson Trevor S , Piersall Savanna L , Lowe Devin B
Published: December 2022

Colorectal cancer remains the third leading cause of cancer-related death in the United States despite advances in targeted therapies such as monoclonal antibodies and tyrosine kinase inhibitors. Progression of vascularized tumors, including colorectal cancer, is reliant on a functioning blood system that provides cancer cells sufficient nutrients and oxygen. This process is ultimately driven by a concerted effort of pro-angiogenic factors, namely vascular endothelial growth factors (VEGFs) and their cognate receptors – the VEGF receptors (VEGFRs). However, tumor vascularization tends to be an unregulated process that results in the overexpression of angiogenic molecules that promote the development of abnormal and chaotic blood vessels, which ultimately impede the perfusion and efficacy of small molecule drugs or immune-based approaches that incorporate cytotoxic CD8+ T cells. FDA-approved targeted agents have been developed against major angiogenic components such as VEGF and VEGFRs, but, despite short-term clinical benefits, these drugs are generally cytostatic and quickly lead to resistance in most patients. To address such shortcomings, active immunotherapeutic strategies directed at auxiliary targets within the tumor vasculature represent a potentially improved direction toward prolonging cancer protection by instituting the long-term destruction of endothelial cell and pericyte networks that provide tumors critical support. We hypothesize that therapeutic vaccination using autologous alpha-type-1 polarized dendritic cells (DCs) pulsed with endothelial cell-and pericyte-derived peptides will induce a protective and safe cytotoxic T cell response in animals. This clinically-relevant DC approach should ultimately lead to colon cancer regressions in mice and allow our group to identify immunologic-based mechanisms that contribute to lasting immunity against the disease.

Citation Format: Amanda L Wooster, Trevor S Anderson, Savanna L Piersall, Devin B Lowe. Analysis of the effects of therapeutic vaccination with tumor vascular-primed dendritic cells in a mouse model of colon cancer [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy; 2022 Oct 21-24; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(12 Suppl):Abstract nr A43.
Abstract A44: Therapeutic blocking of VEGF binding to Neuropilin-2 diminishes PD-L1 expression to activate anti-tumor immunity in prostate cancer
Authors: Wang Mengdie , Wisniewski Christi A , Xiong Choua , Chhoy Peter , Goel Hira , Xu Zhiwen , Loda Massimo , Jiang Zhong , Brehm Michael , Mercurio Arthur
Published: December 2022

Prostate cancers (PC) are largely unresponsive to immune checkpoint inhibitors and there is strong evidence that PD-L1 expression itself must be inhibited to activate anti-tumor immunity. Here, we report that neuropilin-2 (NRP2), which functions as a VEGF receptor on tumor cells, is an attractive target to activate anti-tumor immunity in prostate cancer because we demonstrate that VEGF/NRP2 signaling sustains PD-L1 expression. NRP2 depletion increased T cell activation in vitro and immune-mediated tumor elimination in vivo using a humanized mouse model. Inhibition of the binding of VEGF to NRP2 using a mouse specific anti-NRP2 mAb in a syngeneic model of prostate cancer that is resistant to checkpoint inhibition resulted in significant necrosis and tumor regression compared to control IgG. This therapy also decreased tumor PD-L1 expression and increased immune cell infiltration. We also observed that the NRP2 and VEGF-A genes are amplified in metastatic castration resistant and neuroendocrine prostate cancer (NEPC) and that NRP2high PD-L1high population in metastatic tumors had a significantly lower AR and higher NEPC scores than other populations. Therapeutic inhibition of VEGF binding to human NRP2 with a high affinity humanized mAb, which is suitable for clinical use, in organoids derived from NEPC patients also diminished PD-L1 expression and caused a significant increase in immune-mediated tumor cell killing consistent with the animal studies. These findings provide justification for the initiation of clinical trials using this novel function-blocking NRP2 mAb in prostate cancer, especially for patients with aggressive primary and metastatic cancers, where blocking NRP2-VEGF signaling shows potential in mitigating the morbidity and mortality associated with these cancers.

Citation Format: Mengdie Wang, Christi A Wisniewski, Choua Xiong, Peter Chhoy, Hira Goel, Zhiwen Xu, Massimo Loda, Zhong Jiang, Michael Brehm, Arthur Mercurio. Therapeutic blocking of VEGF binding to Neuropilin-2 diminishes PD-L1 expression to activate anti-tumor immunity in prostate cancer [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy; 2022 Oct 21-24; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(12 Suppl):Abstract nr A44.
Abstract B16: Verteporfin synergizes antitumoral effect of anti-PD-1 in mice cholangiocarcinoma models
Authors: Lee Jihye , Bian Jing , Wang Xin , Cam Meggie , Xie Changqing
Published: December 2022

Cholangiocarcinoma (CCA) is a heterogenous cancer originated from the biliary epithelium in the biliary tree system. The incidence of CCA has gradually increased across nation/worldwide and overall prognosis is poor with only 10% of 5-year survival rate. Recently developed immune checkpoint inhibitor-based immunotherapy has less benefit on CCA patients. We investigated antitumoral effect of verteporfin in combination of anti-PD-1 in YAP/Akt and Notch/Akt hydrodynamic injected mouse model. Our result showed reduced liver weight in verteporfin treated mice group compared to vehicle treated mice group, both in anti-PD-1 and isotype control groups. Single cell RNA sequence showed there is reduced expression of stemness related cells from data observed in verteporfin treated group. B cell, CD8+ T cell, TAM M1 populations were increased in verteporfin treatment group and similar pattern was observed in anti-PD1 treated verteporfin group. In conclusion, verteporfin reduced tumorigenesis in CCA via control of cancer stemness in CCA animal models with anti-PD-1 immunotherapy.

Citation Format: Jihye Lee, Jing Bian, Xin Wang, Meggie Cam, Changqing Xie. Verteporfin synergizes antitumoral effect of anti-PD-1 in mice cholangiocarcinoma models [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy; 2022 Oct 21-24; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(12 Suppl):Abstract nr B16.
Abstract B21: Elucidating the molecular mechanism of Myeloid Derived Suppressor Cell (MDSC) divergence in the tumor microenvironment (TME) of pancreatic cancer
Authors: Dahal Ankit , Hong Yeonsun , Kim Minsoo
Published: December 2022

Pancreatic cancer is one of the deadliest malignancies with an average five-year survival of mere 2-9%. Emerging evidence suggests that the presence of polymorphonuclear (PMN) myeloid derived suppressor cells (MDSCs) with neutrophil-like phenotypes and morphologies play a major role in adverse clinical outcomes due in part to their contribution in establishing immunosuppressive tumor microenvironment (TME). Despite recent advances in our understanding of MDSC phenotypes and characteristics in the TME, the underlying molecular mechanisms by which classical neutrophils differentiate into MDSC within the TME remails largely unknown. To investigate the microenvironmental cues that regulate PMN-MDSC differentiation at the tumor site, we established several in vitro co-culture systems using pancreatic cancer cells and 3D spheroid conditions. With live cell migration microscopy, we first showed that neutrophils actively swarmed into the pancreatic cancer spheroids and stayed active for an extended time period in the tumor. We further found that cancer cell culture supernatant significantly extended neutrophil survival and successfully differentiated neutrophils into MDSCs that exhibited decreased type 1 Interferon levels, increased arginase and ROS expression, and had the ability to suppress CD8 cytotoxicity. Additionally, when compared to other forms of neutrophil activators such as LPS and TNF, only tumor conditioned media enhanced neutrophil survival and MDSC differentiation. Our data suggest the presence of novel molecular processes that support MDSC differentiation at the tumor site. To identify the molecular regulators of MDSC formation, we have developed a CRISPR-Cas9 pooled library screen using the immortalized granulocyte-monocyte progenitor cell line, HoxB8. This study will further our understanding of MDSC differentiation and may help to develop therapeutics to limit MDSC development in the TME of pancreatic and other solid cancers.

Citation Format: Ankit Dahal, Yeonsun Hong, Minsoo Kim. Elucidating the molecular mechanism of Myeloid Derived Suppressor Cell (MDSC) divergence in the tumor microenvironment (TME) of pancreatic cancer [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy; 2022 Oct 21-24; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(12 Suppl):Abstract nr B21.
Abstract B24: Development of targeted T-cell cancer immunotherapies based on a novel enantiomeric cationic lipid that promotes antigen cross-presentation and upregulation of type I interferons
Authors: Gandhapudi Siva K , Sundarapandiyan Karuna , Ward Martin , Fatima Afsheen , Dorrani Mania , Banoub Mary , Dervan Joe , Wood Lauren , Conn Greg , Woodward Jerold G
Published: December 2022

The enantiomeric cationic lipid R-DOTAP nanoparticle platform (Versamune®) was reported (Gandhapudi et. al. 2019, J. Immunol.) to promote cross presentation of multi-epitope human papilloma virus (HPV) antigens and upregulation of Type I interferons, leading to induction of high levels of antigen-specific cytolytic polyfuctional CD8+ T-cells in vivo and complete regression of TC-1 tumors in preclinical models. In two ongoing Phase 2 human clinical trials (ASCO 2022), the early data in a group of the first 60 patients receiving PDS0101 (Versamune® plus HPV16 peptide mix) and who had previously failed standard of care suggests translation of the established mechanism of action into humans resulting in the induction of tumor-specific, clinically effective immune responses. In refractory and difficult-to-treat HPV-related cancers, improved patient survival and tumor shrinkage has been demonstrated. In this study we demonstrate expansion of the Versamune® platform to the treatment of non-viral associated cancers. We have developed Versamune® based drug formulations containing novel multi-epitope peptide sequences of the TARP (T-Cell receptor gamma chain Alternate Reading frame Protein) antigen expressed in 100% of adult and pediatric AML (acute myelogenous leukemia), over 90% of prostate cancers and over 50% of breast cancers. We have also developed Versamune® based drug formulations containing modified sequences of novel Mucin 1 oncoprotein (MUC1) agonist peptide antigens that are over expressed in multiple solid tumors. Using humanized HLA-A2 transgenic mice, we show that the Versamune® platform similarly promotes cross presentation of the non-viral antigens resulting in equally high levels of antigen-specific CD8+ T-cells as observed with HPV. These data demonstrate superior immunogenicity in breadth and potency of vaccine-induced immune responses, supporting progression of the targeted T-cell immunotherapies into human clinical trials.

Citation Format: Siva K Gandhapudi, Karuna Sundarapandiyan, Martin Ward, Afsheen Fatima, Mania Dorrani, Mary Banoub, Joe Dervan, Lauren Wood, Greg Conn, Jerold G Woodward. Development of targeted T-cell cancer immunotherapies based on a novel enantiomeric cationic lipid that promotes antigen cross-presentation and upregulation of type I interferons [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy; 2022 Oct 21-24; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(12 Suppl):Abstract nr B24.
Abstract B38: Single-cell RNA-seq analysis of a soft-tissue sarcoma model reveals the critical role of tumor-expressed MIF in shaping macrophage heterogeneity
Authors: Guarnerio Jlenia
Published: December 2022

Soft-tissue sarcomas (STS) are mesenchymal tumors that affect almost 200,000 individuals worldwide each year. Surgery is the standard treatment, followed by anthracycline-based chemotherapy, either alone or in combination with ifosfamide. Still, the 5-year survival rate is below 20% for advanced and metastatic STS. A main clinical challenge is creating and implementing therapies that can successfully treat the primary tumors so that the disease burden is eradicated, and relapsing lesions are prevented. Immunotherapy approaches such as anti-PD1/PDL1 have produced only marginal results for sarcoma patients. Novel investigations in this respect are hindered by the lack of knowledge of the sarcoma immune microenvironment and the scarcity of STS immune-competent mouse models. We therefore developed a mouse model of undifferentiated pleiomorphic sarcoma (UPS), one of the most aggressive STS subtypes, and employed single-cell RNA-sequencing to characterize the immune composition of the UPS mass. scRNA-seq showed that macrophages, the main immune cells in sarcoma, exhibit distinct activation states, ranging from anti-tumorigenic to pro-tumorigenic. Notably, sarcoma cells used the pleiotropic cytokine Macrophage Migration Inhibitory Factor (MIF) to interact with macrophages expressing the CD74 receptor, to switch macrophages’ activation state and pro-tumorigenic potential. Blocking the expression of MIF in sarcoma cells favored the accumulation of macrophages with inflammatory and antigen-presenting profiles, hence reducing tumor growth. These data pave the way for testing new therapies aimed at re-shaping the sarcoma microenvironment, in combination with the standard of care.

Citation Format: Jlenia Guarnerio. Single-cell RNA-seq analysis of a soft-tissue sarcoma model reveals the critical role of tumor-expressed MIF in shaping macrophage heterogeneity [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy; 2022 Oct 21-24; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(12 Suppl):Abstract nr B38.
Abstract B41: Reprogramming immunosuppressive tumor-associated macrophages potentiates standard-of-care therapy in melanoma
Authors: Tichet Melanie , Chryplewicz Agnieszka , Hanahan Douglas
Published: December 2022

Cutaneous melanoma is a highly aggressive cancer capable of distant and lethal metastatic spread. Recent breakthroughs have come from understanding oncogenic signaling and cancer immunobiology. Targeted therapies successfully block MAPK signaling in BRAFV600e mutant melanoma with remarkably high clinical responses followed by rapid relapse, whereas checkpoint inhibitors activating the immune response induce long-lasting responses, albeit only in a subset of patients. These limitations have driven interest in understanding innate and acquired resistance. Using an immunocompetent genetically-engineered mouse model of BRAF-driven melanoma, which phenocopies the human disease in its development, histopathology, and response to therapy, we focused on the tumor microenvironment (TME), seeking to elucidate resistance mechanisms. Our investigations have revealed that tumor-associated macrophages (TAMs) are a critical component of the TME, predominantly polarized toward a pro-tumoral “M2-like” phenotype, producing immunosuppressive factors and exhibiting extensive immuno-suppressive capabilities suggesting that they might significantly hinder immune responses in the melanoma TME. Seeking to assess their functional importance, we have found that combining conventional strategies with TAMs-reprogramming agents stimulates anti-tumor immune responses, leading to improved survival and responsiveness to standard-of-care therapies. These data highlight the central role played by macrophages in melanoma progression and demonstrate that pharmacologic reprogramming of macrophages represents a new therapeutic modality with the potential to elicit more effective anti-tumor immune responses against this devastating disease.

Citation Format: Melanie Tichet, Agnieszka Chryplewicz, Douglas Hanahan. Reprogramming immunosuppressive tumor-associated macrophages potentiates standard-of-care therapy in melanoma [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy; 2022 Oct 21-24; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(12 Suppl):Abstract nr B41.