bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1129: A multifaceted study of X-Ray radiation therapy across diverse mouse tumor models
Authors: Huang Jingqi , Li Wentao , He Xiaoyan , Cheng Weiwei , Zhang Lingyun , Li Guannan , Li Qiuliang , Wang Yongfei , Ren Xuesong , Wei Zhi , Shi Long , Wei Yiran , Jin Jing , Li Linfeng , Yun Wei
Published: March 2024

In this study, we showcase the application of X-ray radiation therapy of tumor-bearing mice across 13 syngeneic tumors, one intracranial tumor, and in combination with radiosensitizing drugs, aiming to develop potent cancer treatment strategies. 1.We examined the effects of different radiation levels on multiple tumor models using a therapeutic device delivering targeted radiation to the tumor site. 2.We investigated the combined benefits of radiation with the radiosensitizer Gemcitabine on the H22 liver tumor model. 3.We assessed the impact of radiation sensitization on the HCC1975-luc intracranial tumor model in combination with AZD0156, a drug that cannot cross the blood-brain barrier. The integrity of the blood-brain barrier and the presence of the pharmacodynamic marker pRAD50 of AZD0156 were evaluated. The results show that X-ray radiation has anti-tumor effects across diverse models, with combination drug treatment with both Gemcitabine and AZD0156 showing enhanced therapeutic effects. Furthermore, pRAD50 showed decreasing trend in the single dose PD study in HCC1975-luc intracranial model. We conclude that our platform provides robust methods for evaluating the therapeutic effects of X-Ray radiation, offering invaluable insights for the creation of new cancer therapies.

Citation Format: Jingqi Huang, Wentao Li, Xiaoyan He, Weiwei Cheng, Lingyun Zhang, Guannan Li, Qiuliang Li, Yongfei Wang, Xuesong Ren, Zhi Wei, Long Shi, Yiran Wei, Jing Jin, Linfeng Li, Wei Yun. A multifaceted study of X-Ray radiation therapy across diverse mouse tumor 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 1129.
Abstract 113: Generation and validation of a novel humanized IL-31/IL-31RA/OSM/OSMR mouse model
Authors: Wang Linlin , Zhao Lei , Zhang Meiqi , Zhang Zhi , Chen Zhen , Kuraguchi Mari , Lian Jingjing , Zhang Jiansu
Published: March 2024

Oncostatin M (OSM) and interleukin-31 (IL-31) are two pleiotropic cytokines that share a common signaling receptor subunit, the OSM receptor beta (OSMR). Both cytokines are released by monocytes, macrophages, dendritic cells and T lymphocytes in inflammatory situations, and upon binding to their respective receptors, induce the production of inflammatory cytokines (such as IL-6), chemokines, and other cellular growth factors during tissue repair, mainly via the JAK/STAT signal pathway. As OSM and IL-31 are reported to be involved in a variety of physiological and pathological processes, targeting IL-31 or OSM signaling is of therapeutic interest. To provide a novel preclinical platform for testing novel modulators of these signaling axes, a humanized IL-31/IL-31RA/OSM/OSMR mouse model was generated. First, single gene knock-in mice were constructed by gene editing technology; multigenic knock-in mice were then obtained by breeding, yielding a quadruple humanized model in which the murine cytokine sequences were replaced by full-length human cytokine sequences in situ, while the murine receptor sequences were replaced by chimeric sequences comprised of the human extracellular region and murine intracellular regions in situ. mRNA expression confirmed expression of IL-31, IL-31RA, and OSMR in this model, while ELISA confirmed expression of human OSM protein in the model without evidence of murine OSM expression. Next, the signaling capacity of the humanized OSMR receptor was confirmed: mouse embryonic fibroblasts (MEFs) isolated from humanized B-hOSM/hOSMR mice were capable of responding to human OSM to release IL-6, while wild-type MEFs could only respond to murine OSM. Subsequently, the ability of IL-31RA antibodies to alleviate scratching was tested in a chemically induced model of atopic dermatitis. Compared to mice receiving control antibodies, scratching frequency in humanized B-hIL31/hIL31RA/hOSMR mice treated with nemolizumab was attenuated (analog antibody was generated in-house). Finally, blood counts and flow cytometric profiling indicates that the humanized mice have a normal frequency of leukocytes and lymphocytes. Taken together, this quadruple humanized IL-31/IL-31RA/OSM/OSMR mouse model is a promising preclinical tool that can be used to evaluate novel modulators of this signaling pathway prior to clinical development.

Citation Format: Linlin Wang, Lei Zhao, Meiqi Zhang, Zhi Zhang, Zhen Chen, Mari Kuraguchi, Jingjing Lian, Jiansu Zhang. Generation and validation of a novel humanized IL-31/IL-31RA/OSM/OSMR 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 113.
Abstract 1161: Deep spatial proteomics: A new approach for obtaining insight into the glioblastoma microenvironment
Authors: Saxena Kartikey , Daucke Rune , Pedersen Vilde , Klausen Pia Helene , Schoof Erwin , Kristensen Bjarne Winther
Published: March 2024

Glioblastoma is the most frequent and malignant brain tumor, with a median survival of approximately 14 months. The tumor microenvironment is composed of different cell types and is known to support the glioblastoma cells and contribute to poor patient prognosis. Since accumulating studies report discordance between mRNA and protein abundances both on population and single-cell levels, and since proteins are the effector molecules of the cells and the actual target molecules of most drugs, our aim is to dissect the tumor microenvironment by spatial LC-MS based proteomics. Glioblastoma formalin-fixed paraffin-embedded (FFPE) tissue was sectioned and placed on membrane glass slides suitable for laser capture microdissection (LCM), followed by precise isolation of single cells from the heterogeneous tumor microenvironment. We used the Zeiss Palm MicroBeam laser microdissection instrument, combined with an Orbitrap Eclipse Tribrid Mass Spectrometer running our in-house sensitivity tailored Data Independent Acquisition method (WISH-DIA) to analyze and quantify the proteomes. We obtained up to 3000 proteins out of only 100 cells and were able to demonstrate the feasibility of collecting and analyzing various cell types at the single-cell level. The proteins identified belonged to many different classes/families of proteins and were able to distinguish the different cell types. In conclusion, this on-going study demonstrates our ability to isolate single cells from the highly heterogeneous tumor microenvironment of glioblastoma with ultra-high sensitivity, thereby allowing the characterization of the proteome of the glioblastoma microenvironment followed by identification of potential therapeutic and/or diagnostic targets.

Citation Format: Kartikey Saxena, Rune Daucke, Vilde Pedersen, Pia Helene Klausen, Erwin Schoof, Bjarne Winther Kristensen. Deep spatial proteomics: A new approach for obtaining insight into the glioblastoma microenvironment [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 1161.
Abstract 1244: Exploiting the pathogenesis of endogenous retrovirus to tackle squamous cancers
Authors: Lyu Ying , Ge Yejing
Published: March 2024

Retrotransposons are the evolution relic of ancestrally invaded retroviruses interspersed through our genome. Mostly domesticated as host regulome, retrotransposons shape lineage gene expressions or donate coding functions. A few rare evolutionarily young species remain virally active, leading to germline mutations and modulating immune functions. Aberrant retrotransposon activities have been widely observed in adult diseases, although the extent to which they ensue active viral form and function under those contexts is unclear. Their molecular trigger in adult pathophysiology is poorly defined. To address these questions, we use murine skin as our model, given its well-characterized, abundant and highly accessible adult stem cells, not only mediating postnatal remodeling and homeostatic regeneration, but also responding to stress by driving tissue repair and adaptations. By analyzing a genetic model lacking a known retrotransposon suppressor, histone methyltransferase Setdb1, which we found to be essential in the adult skin, we saw hair loss and hair follicle stem cell exhaustion phenotype, accompanied by a robust and selective surge of endogenous retroviruses (ERVs). When combined with squamous cell carcinoma drivers, Setdb1 loss significantly blocked tumor progression in vivo. Interestingly, compared to relatively mild or no induction of commonly observed evolutionarily young ERVs, the most induced species in our model is a class I ERV, murine leukemia virus (MuLV). We detected abundant retroviral peptides originated from its full-length copies through mass spectrometry and viral-like particles through transmission electron microscopy that are immunoreactive to a monoclonal MuLV env antibody. Similar viral-like particles were broadly evident across several epithelial tissues beyond skin, implicating its conserved function across squamous cancers. Epithelial originated ERVs elicit tissue wide inflammatory response, and can be ameliorated by pharmacological or genetic inhibition of retroviral activity. Physiologically, viral-coding ERVs arise as the epigenetic machinery declines in response to age-associated stress, and plummet when host safeguard prevails in UV induced skin hyperplasia. As an evolutionary conundrum, viral-coding ERVs pose a threat to host fitness and yet, they’ve resisted extinction persisting in the mammalian genome. Our findings suggest viral-coding ERVs wrestle with the host surveillance programs to regulate adult tissue physiology and pathology, thus providing a key target in eliciting innate immunity and tackling squamous cancers.

Citation Format: Ying Lyu, Yejing Ge. Exploiting the pathogenesis of endogenous retrovirus to tackle squamous cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1244.
Abstract 1275: In vivo functional CRISPR screens identifies metabolic dependencies mediating triple negative breast cancer lung metastasis
Authors: Wang Xiaoyong , Chou Shu-ting , Ngwa Verra , Hwang Yoonha , Karno Breelyn , Edwards Deanna , Chen Jin
Published: March 2024

Metastasis is responsible for about 90% of cancer-related mortality. Throughout the metastatic cascade, cancer cells requires substantial metabolic rewiring to adapt the nutrient micro-environmental changes enabling dissemination and seeding in distant organs. Recent insights into importance of the tumor-intrinsic metabolic reprograming has been implicated in immune escape, yet altered immune responses imposed by tumor metabolism during metastasis are poorly understood. Here, we generated genetic perturbations in triple-negative breast cancer cells using metabolism-focused CRISPR knockout library and performed in vivo screens in both immunocompetent and immunodeficient mice. By comparing enriched sgRNAs in lung metastases or primary tumors, we identify loss of one gene related to long chain fatty acid (LCFA) β-oxidation (FAO) potentiates metastatic dissemination in immunocompetent mice but no in immunodeficient mice, suggesting that impaired FAO in tumor cells promotes breast cancer lung metastasis through an lymphocytes-dependent manner. We confirmed individual sgRNAs targeting this FAO gene from the primary screen dramatically accelerate metastasis in different breast cancer orthotopic models. Specifically, we find that low expression of this gene in primary tumors of advanced breast cancer patients with high grade tumors and positive lymph nodes (LN+) is associated with reduced overall survival. In addition, deletion of this gene generates an immunosuppressive microenvironment in the lung metastatic tumors with increased neutrophils, monocytes and decreased TNFα+-IFNγ+ CD8+ T cells. Mechanistically, impaired FAO in tumor cells causes mitochondria imbalance under lipid metabolism stress, consequently triggers inflammatory response on establishing a neutrophil-rich metastatic niche via mitochondrial nucleic acid-driven innate immunity. Altogether, our data provides a new insight on how tumor cell lipid metabolism impacting immune interplay during metastatic progression, nominating potential anti-tumor immunotherapies.

Citation Format: Xiaoyong Wang, Shu-ting Chou, Verra Ngwa, Yoonha Hwang, Breelyn Karno, Deanna Edwards, Jin Chen. In vivo functional CRISPR screens identifies metabolic dependencies mediating triple negative breast cancer lung metastasis [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 1275.
Abstract 1276: Kindlin-2: a novel target of Parkin regulating cancer metastasis
Authors: Yeon Minjeong , Bertolini Irene , Altieri Dario C.
Published: March 2024

Although targeted therapies in prostate cancer have clinically shown successful outcomes, resistance to these therapies and acquisition of lineage plasticity pose challenges for fully treating prostate cancer. Among several microscopic perspectives to solve these obstacles, much evidence has emerged indicating that mitochondria dynamics are closely associated with tumor metastasis and tumor progression in various types of cancer.

The E3 ubiquitin ligase Parkin is associated with PINK1-mediated mitophagy, allowing cells to control mitochondria damages and generate the needed energy to survive in harsh physiological conditions.

Interestingly, we found that Parkin potentially ubiquitinates proteins independently of PINK1-associated mitophagy. By SALIC analysis, we found that K681 and K582 of Kindlin-2 is one of the potential ubiquitination sites by Parkin in prostate cancer cells.

Kindlin-2 is a cytoskeletal membrane protein and is a component of the focal adhesion complex. Computational modeling suggested that N-terminal region of Kindlin-2 binds to Parkin, with K581 and K582 of Kindlin-2 exposed outside for ubiquitination. Our studies showed that Kindlin-2 is ubiquitinated and degraded by wild-type Parkin but not by the catalytic site, C431S, mutant Parkin. Furthermore, K581A and K582A mutant Kindlin-2 did not exhibit poly-ubiquitination under conditions of Parkin overexpression. Surprisingly, in our immunofluorescence study, Kindlin-2 was recruited to the mitochondria and co-localized with Parkin. From a mitochondrial functional perspective, wild-type Kindlin-2 reduced mitochondria dynamics, whereas mutant Kindlin-2 didn’t in the presence of Parkin. This suggests that the cytoskeletal protein Kindlin-2 might play a role in mitochondrial dynamics that mediates tumor metastasis.

Wild-type Kindlin-2 reduced both focal adhesion generation and decay, unlike mutant Kindlin-2 under Parkin-induced condition. Additionally, this pattern was observed in the single cell lamellipodia dynamics through live cell imaging. 2D and 3D cell invasive assays supported the evidence that Parkin decreases tumor metastatic potential by ubiquitinating Kindlin-2 and regulating mitochondria and focal adhesion dynamics.

Our findings indicate that Parkin reduces tumor metastatic potential by ubiquitinating Kindlin-2 and demonstrate the possibility as a therapeutic agent for advanced and immune suppressive tumors. Furthermore, Parkin could be a promising agent for Parkin suppressed tumor patients in line with personalized cancer therapy development.

Citation Format: Minjeong Yeon, Irene Bertolini, Dario C. Altieri. Kindlin-2: a novel target of Parkin regulating cancer metastasis [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 1276.
Abstract 1334: Evaluation of NK cell combination therapies in mouse models of cancer
Authors: Franciszkiewicz Katarzyna , Rettman Pauline , Gauthier Beatrice , Noblet Laurence , Nicolazzi Celine , Kunert Andre , Igarashi Rob , Calvet Loreley , Fantin Valeria , Sidhu Sukhvinder
Published: March 2024

Adoptive immunotherapy with natural killer (NK) cells has emerged as a promising innovative approach against hematological malignancies and solid tumors. To enhance the functional aspects of NK cells and to achieve stronger, sustained anti-tumor responses, combination strategies are under development. They are focused on optimization of NK cell potency, persistence and tumor targeting using monoclonal antibodies, multi-specific NK cell engagers and immune modulators like cytokines. SAR445419 is a universal off-the-shelf allogeneic human NK cell (hNK) product made using Sanofi’s proprietary platform with potential broad application across hematologic and solid tumor indications. We evaluated the therapeutic potential of these NK cells in combination with internal assets and standard of care treatments in preclinical mouse models. The studies were performed in immunodeficient NOG mouse strains transgenic for hIL-15, supporting long-term engraftment of functional hNK cells. We demonstrated the therapeutic advantage of NK cell combination with an anti-CD20 monoclonal antibody (mAb) in a disseminated lymphoma model. We also reported prolonged lifespan of leukemia-bearing animals treated with NK cells and pegenzileukin, a PEGylated non-alpha rhIL-2 variant engineered to preferentially bind to the IL-2 βγ receptors and promote NK cell function. The anti-tumor response correlated with pegenzileukin-induced increase in NK cell numbers in the blood. A combination of CD38KO NK cells with an anti-CD38 mAb was also investigated in a model of multiple myeloma (MM). FcγR-KO immunodeficient mice were used in these studies to limit the confounding contribution of residual murine immune cells engaging with the Fc-competent therapeutic antibody and to specifically detect hNK-cell mediated activity. The comparison of CD38KO versus WT NK cell numbers in the blood of animals treated with an anti-CD38 mAb showed a significant decrease in CD38-expressing NK cell count that was mitigated by the genetic deletion of CD38. Obtained results suggest that CD38KO NK cells are resistant to anti-CD38 mAb-mediated fratricide and provide a rationale for the combination of CD38KO NK cells with anti-an CD38 mAb against MM. Overall, these preclinical in vivo studies have a pivotal role in the evaluation of NK cell combination therapies against cancer and reveal their therapeutic potential.

Citation Format: Katarzyna Franciszkiewicz, Pauline Rettman, Beatrice Gauthier, Laurence Noblet, Celine Nicolazzi, Andre Kunert, Rob Igarashi, Loreley Calvet, Valeria Fantin, Sukhvinder Sidhu. Evaluation of NK cell combination therapies in mouse models of 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 1334.
Abstract 134: ZEB2 as a therapeutic target in fusion-positive rhabdomyosarcoma
Authors: Fahs Assil , Munshi Aanandi , Barbar Leen , Zimmerman Seth , Counter Chris , Linardic Corinne
Published: March 2024

Rhabdomyosarcoma (RMS) is a highly aggressive childhood soft tissue cancer with skeletal muscle histogenesis. It is classified at the molecular level into fusion-negative (FN) and fusion-positive (FP) subtypes, with FP subtypes being defined by their signature oncofusions between the transcription factors PAX3 (or PAX7) and a variety of 3’ partners. Patients with FP-RMS have a five-year survival of <30%, and current treatments remain ineffective against the driving oncofusions. To address this challenge, we are interested in identifying proteins that interact with the oncofusions as alternate therapeutic targets, and conducted an unbiased proximity labeling screen for seven of the known RMS oncofusions. This approach revealed that ZEB2 is a common member of the interactome for all seven FP-RMS subtypes. ZEB2 is a DNA-binding transcriptional repressor that interacts with SMAD proteins and is known to be important for coordinating muscle and nerve development during embryogenesis. ZEB2 has been identified as an oncogene in various cancer types such as leukemia, glioblastoma, lung, and breast cancers. Further analysis of ZEB2's expression in FP-RMS cell lines, compared to skeletal muscle and other cancer cell lines, showed significantly higher levels of expression. In silico analyses of genomic CRISPR and RNAi screens in DepMap suggested a strong dependency of FP-RMS on ZEB2, with ZEB2 being one of the top four differential CRISPR dependencies in RMS cell lines compared to all other cancer cell lines. To investigate the functional consequences of ZEB2 suppression, we employed a loss-of-function approach using RNAi (siRNA and shRNA) in human FP-RMS cells. Our experiments confirmed the knockdown of ZEB2 through qRT-PCR and immunoblotting. Interestingly, the extent of ZEB2 knockdown determined whether FP-RMS cells underwent apoptosis or differentiation, potentially towards a neurodifferentiation pathway. We hypothesize that ZEB2 holds potential as a therapeutic target for FP-RMS patients. We aim to elucidate ZEB2's phenotypic role and mechanism of action in FP-RMS tumorigenesis, and assess its value as a therapeutic target in vivo. Our long-term goals for this project are to contribute to the development of innovative therapies that can mitigate the oncogenic activity driven by oncofusions in FP-RMS.

Citation Format: Assil Fahs, Aanandi Munshi, Leen Barbar, Seth Zimmerman, Chris Counter, Corinne Linardic. ZEB2 as a therapeutic target in fusion-positive 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 134.
Abstract 1344: Improvement of the anti-tumor activity of NK cells and the efficacy of anti-CD20 monoclonal antibodies in the therapy of B cell-derived malignancies
Authors: Zerrouqi Abdessamad , Zdanowicz Aleksandra , Torun Anna , Ciechanowicz Andrzej , Pradhan Bhaskar , Rymkiewicz Grzegorz , Efremov Dimitar , Pyrzynska Beata
Published: March 2024

Resistance of tumor cells to immunotherapy with monoclonal antibodies (mAbs) often relies on the reduced level of the antigen targeted by the particular mAb. We have therefore screened for the compounds able to upregulate selected antigens on the surface of lymphoma/leukemia cells, to minimize the possibility of resistance acquired by the tumor cells. We have found that members of a specific class of antibiotics, known as cation carriers, namely salinomycin (SAL), monensin (MON), narasin, and nigericin exhibit great CD20 antigen-upregulating abilities. A variety of Burkitt’s lymphoma and diffuse large B-cell lymphoma (DLBCL) cell lines as well as patients-derived primary malignant cells, such as DLBCL and chronic lymphocytic leukemia (CLL) cells exhibited upregulation of surface CD20 upon the treatment with SAL and MON at concentrations as low as 250nM and 50 nM, respectively. As a consequence, these compounds appeared to be beneficial for the therapy of lymphoma/leukemia with anti-CD20 mAbs, both in vitro and in animal models.Moreover, the proteomics analysis revealed that besides CD20, cation carriers upregulated a number of different proteins on the surface of tumor cells, including proteins implicated in the interaction with immune cells. These included ICAM1, HLA-A/B/C, CD155, CD80, CD86 and ULBP2/5/6. Particularly important appears to be the upregulation of antigens interacting with the activating receptors on NK cells, helping them to overcome the inhibitory signals and stimulate the lysis of tumor cells. Thus, the results of proteomics analysis explained why SAL- or MON-pretreated tumor cells were efficiently eliminated by NK cells, even in the absence of therapeutic mAb. In light of our discoveries, it seems reasonable to consider the cation carriers as promising drug candidates for the improvement of NK cell anti-tumor activity and potential treatment of B-cell non-Hodgkin lymphoma patients, in combination with anti-CD20 mAbs.Funding: National Science Centre (NCN, Poland), grants 2019/35/B/NZ5/01445 and 2020/39/B/NZ6/03513.

Citation Format: Abdessamad Zerrouqi, Aleksandra Zdanowicz, Anna Torun, Andrzej Ciechanowicz, Bhaskar Pradhan, Grzegorz Rymkiewicz, Dimitar Efremov, Beata Pyrzynska. Improvement of the anti-tumor activity of NK cells and the efficacy of anti-CD20 monoclonal antibodies in the therapy of B cell-derived malignancies [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 1344.
Abstract 1358: Elucidating the contribution of T cell-intrinsic VISTA to the suppression of antitumor T cell immunity
Authors: Patnaik Sachin , Delaney Elizabeth , Gilmour Cassandra , Zhang Keman , Zakeri Amin , Ta Hieu M. , Parthasarathy Prerana B. , Alban Tyler , Chan Timothy , Wang Li Lily
Published: March 2024

Immune checkpoint receptors (ICRs) negatively regulate antitumor immune responses. While significant advances have been made in understanding the mechanisms of well-established ICRs such as PD-1 and CTLA-4, other checkpoint proteins remain incompletely characterized and may be targeted to synergistically boost antitumor immunity. One such protein is V-domain immunoglobulin suppressor of T-cell activation (VISTA), an ICR whose expression curtails the function of several lymphoid and myeloid lineages, including tumor-reactive cytotoxic T cells. Globally blocking VISTA elicits potent T cell-mediated antitumor immunity and systemic myeloid cell-mediated inflammatory responses. Mechanistically, VISTA has been shown to inhibit T cell activation by acting as an inhibitory ligand on antigen-presenting cells. However, the impact of T cell-intrinsic VISTA on modulating tumor-reactive T cell responses has not been thoroughly investigated, and it remains unclear whether blocking T cell-specific VISTA is adequate for revitalizing T cell-mediated tumor control. In this study, we utilized VISTA conditional knockout mice models to examine the relative contribution of T cell-intrinsic VISTA versus myeloid-derived VISTA in the regulation of antitumor immunity. We present evidence that blocking VISTA on T cells alone is insufficient to induce strong antitumor immune responses, whereas blocking VISTA on myeloid cells counteracts its dominant role in orchestrating T cell suppression.

Citation Format: Sachin Patnaik, Elizabeth Delaney, Cassandra Gilmour, Keman Zhang, Amin Zakeri, Hieu M. Ta, Prerana B. Parthasarathy, Tyler Alban, Timothy Chan, Li Lily Wang. Elucidating the contribution of T cell-intrinsic VISTA to the suppression of antitumor T cell immunity [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 1358.
Abstract 1395: Unraveling the interplay between tumor heterogeneity and microenvironment in pancreatic ductal adenocarcinoma evolution and its clinical implications
Authors: Kim Hyemin , Jeong Hyong-oh , Lee Se-Hoon , Lee Kwang Hyuck , Lee Kyu Taek , Lee Jong Kyun , Lee Min Woo , Lee Semin , Park Joo Kyung
Published: March 2024

Intratumoral heterogeneity (ITH) and tumor microenvironment (TME) of pancreatic ductal adenocarcinoma (PDAC) play important roles in tumor evolution and patient outcomes. However, the precise characterization of diverse cell populations and their crosstalk associated with PDAC progression and metastasis is still challenging. We performed single-cell RNA sequencing of primary PDAC samples with and without paired liver metastasis samples to understand the interplay between ITH and TME in the PDAC evolution and its clinical associations. We discovered that even a small proportion (~30%) of basal-like ductal cells in primary PDAC was associated with poor chemotherapy response and patient survival and that epithelial-mesenchymal transition programs were largely subtype specific. The clonal homogeneity significantly increased with more prevalent and pronounced copy number gains of oncogenes, such as KRAS and ETV1, and losses of tumor suppressor genes, such as SMAD2 and MAP2K4, along PDAC progression. Diverse immune cell populations, including naive SELLhi regulatory T cells (Tregs) and activated TIGIThi Tregs, contributed to shaping immunosuppressive TMEs of PDAC through cellular interactions with malignant ductal cells. The proportion of basal-like ductal cells negatively correlated with that of immunoreactive cell populations, such as cytotoxic T cells, but positively correlated with that of immunosuppressive cell populations, such as Tregs. Overall, our study provides a valuable resource for better understanding the roles of ITH and TME in PDAC evolution and their clinical relevance in the treatment of PDAC.

Citation Format: Hyemin Kim, Hyong-oh Jeong, Se-Hoon Lee, Kwang Hyuck Lee, Kyu Taek Lee, Jong Kyun Lee, Min Woo Lee, Semin Lee, Joo Kyung Park. Unraveling the interplay between tumor heterogeneity and microenvironment in pancreatic ductal adenocarcinoma evolution and its clinical implications [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 1395.
Abstract 1405: Development of a humanized anti-IL-22 antibody for cancer and inflammation therapy
Authors: Chen Jian , Chang Cathy , Frey Gerhard , Liu Haizhen , Wang Jing , Boyle William J. , Short Jay M.
Published: March 2024

Interleukin-22 (IL-22) is an inflammatory cytokine involved in the pathology of autoimmune diseases such as psoriasis, atopic dermatitis, and ulcerative colitis. IL-22 has also been shown to promote epithelial cell proliferation, stemness and tumorigenesis in cancer, and is associated with a more aggressive phenotype in a variety of cancers including inflammatory colon cancer models. IL-22 plays a central role in this pathogenic process by driving activation of STAT3 and other signaling cascades. Recent studies demonstrated that neutralization of IL-22 reduced dysplasia and tumor development in preclinical models. However, there has not been a clear application for anti-IL-22 therapy for cancer and autoimmune diseases, likely due to identifying the right disease indication and their low potency and efficacy. Thus, there remains an opportunity to explore the potential of more potent anti-IL-22 therapeutics.

We have developed humanized anti-IL-22 antibodies with high affinities to human, cynomolgus and mouse IL-22 using BioAtla’s proprietary antibody discovery and engineering platforms. In vitro data demonstrated that our anti-IL-22 antibodies inhibited IL-22-induced p-STAT3 activities. In addition, our antibodies showed a 10-fold increased binding activity to human IL-22 compared to Fezakinumab, which was previously developed by Pfizer for the treatment of autoimmune diseases. To test the functions of our antibodies in vivo, we established an inflammation-driven sporadic colitis-associated colorectal (CAC) cancer mouse model. Our humanized anti-IL-22 antibody, a potent, species cross-reactive anti-IL-22 blocking antibody, significantly reduced tumor progression in CAC model. These data suggest that targeting IL-22 reduces tumor occurrence by reducing inflammation-induced tumorigenesis. In addition, we evaluated our anti-IL22 antibodies in an imiquimod-induced psoriasiform skin inflammation mouse model. The mice treated with anti-IL22 antibodies had significantly reduced skin lesions compared to isotype antibody-treated mice. Q-PCR analysis of the mouse skin demonstrated that treatment with our anti-IL-22 antibody led to significantly decreased RNA levels of the imiquimod-induced inflammatory marker CXCL3. In conclusion, the development of a potent, species cross-reactive anti-IL-22 antibody using BioAtla’s antibody discovery and engineering platforms addresses the past challenges of anti-IL-22 therapeutics and allows for translational studies in relevant animal efficacy and safety models.

Citation Format: Jian Chen, Cathy Chang, Gerhard Frey, Haizhen Liu, Jing Wang, William J. Boyle, Jay M. Short. Development of a humanized anti-IL-22 antibody for cancer and inflammation therapy [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 1405.
Abstract 1406: Plk1 attenuates colon inflammation and tumorigenesis
Authors: Wang Xinyi , he Dang , Mao Fengyi , Kong Yifan , Li Chaohao , Wu Sai , Peng Jia , Wang Chi , Li Zhiguo , Liu Xiaoqi
Published: March 2024

Inflammation-associated colon cancer, linked with long-standing colon inflammation, is often seen in patients with inflammatory bowel diseases (IBD), such as Crohn’s disease and ulcerative colitis. Typically, these patients are diagnosed at a younger age compared to those with sporadic colon cancer. Despite some advancements, the detailed molecular mechanisms driving cancer development in IBD remain unclear. Polo-like kinase 1 (Plk1), a serine/threonine kinase and cell cycle regulator, is recognized as an oncogene in various cancers. Intriguingly, higher Plk1 expression correlates with better survival rates in colon cancer patients. Besides, Plk1 also elevates in IBD cases, suggesting its significant role in both colitis and colon cancer. Our study explores Plk1's role in IBD-associated colon cancer using the azoxymethane (AOM)/dextran sulfate sodium (DSS) mouse model, revealing Plk1's suppressive impact on both colitis and related colon cancer.

Citation Format: Xinyi Wang, Dang he, Fengyi Mao, Yifan Kong, Chaohao Li, Sai Wu, Jia Peng, Chi Wang, Zhiguo Li, Xiaoqi Liu. Plk1 attenuates colon inflammation and tumorigenesis [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 1406.
Abstract 1435: Utilization of CLDN18.2 syngeneic mouse models to study chimeric antigen receptor T cells in immunocompetent mice
Authors: Breen Shannon , Carrasco Rosa , McGlinchey Kelly , Janocha Brianna , Zanvit Peter , Ortiz Lorenzo , Clark Benjamin , Toloczko Aleksandra , Cobbold Mark , Moody Gordon , Bosco Emily , Barrett Allison
Published: March 2024

CLDN18.2 expression has been observed in pancreatic, gastric and esophageal cancers, whereas normal tissue expression of CLDN18.2 is mostly restricted to differentiated gastric mucosal epithelial cells. This expression profile renders CLDN18.2 an ideal tumor associated antigen (TAA) for CAR-T targeting and an armored targeted CAR-T, AZD6422, is in clinical development. While CAR-T are beginning to show clinical promise in solid tumors, a consistent challenge is to overcome the immunosuppressive tumor microenvironment (TME) where immune and stromal cells secrete high levels of TGFb. Since the majority of pre-clinical CAR-T efficacy data is generated utilizing immunocompromised mouse models, which lack a proper TME, the team utilized syngeneic xenograft models to circumvent this challenge. ​ In vitro studies showed that CLDN18.2 targeted murine CAR-T cells generated by retroviral transduction induced efficient cytolysis of target positive mouse tumor cells accompanied by cytokine release. To model the clinical scenario, tumor-bearing mice were lymphodepleted with total body irradiation (TBI) to enhance CAR-T engraftment. The dosage of TBI used in these studies was predetermined to have caused leukopenia and minimal impact on tumor growth. We observed substantial tumor control and extended tumor efficacy in mice treated with mCAR-T cells compared to those infused with untransduced T cells or in non-target bearing control tumors in the absence of gross toxicities. ​ This syngeneic mouse model platform will additionally allow us to investigate how mCAR-T cells perform within a fully competent TME in combination with other therapeutic agents targeting cancer cells via altered mechanism of action. ​

Citation Format: Shannon Breen, Rosa Carrasco, Kelly McGlinchey, Brianna Janocha, Peter Zanvit, Lorenzo Ortiz, Benjamin Clark, Aleksandra Toloczko, Mark Cobbold, Gordon Moody, Emily Bosco, Allison Barrett. Utilization of CLDN18.2 syngeneic mouse models to study chimeric antigen receptor T cells in immunocompetent mice [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 1435.
Abstract 1439: Elucidation of the pathogenesis of prostate cancer harboring mutations of epigenetic factor UTX using genetically engineered mice
Authors: MURAMOTO NOBUHITO , Iwasaki Masayuki , Sera Yasuyuki , Ide Hisamitsu , Mukai Shoichiro , Fukushima Tsuyoshi , Horie Shigeo , Honda Hiroaki
Published: March 2024

Introduction and Objective: Histone modifications are important for tissue homeostasis, and their mutations are involved in carcinogenesis. Mutations in a histone modifier, UTX (KDM6A) are common in prostate cancer (PCa), which is frequently associated with loss of its counterpart UTY (KDM6C). However, mechanisms underlying the development of UTX-deficient PCa are not clear. We aimed to address this issue by generating and analyzing genetically-engineered mice.

Methods: We generated mice with deletion of both Utx and Uty in the prostate tissue (UtxΔ, UtyΔ) and crossed with mice with a heterozygous deletion of p53 (p53+/−) to create UtxΔ, UtyΔ and p53+/− compound mice. High-fat diet (HFD) that is known to promote cancer growth was administered to mice. We found that UtxΔ, UtyΔ, p53+/− mice treated with HFD developed PCa with Gleason score (GS) = 3+3. Isolated prostate tissues were subjected to pathologic, molecular, and cellular analyses. In addition, RM-2 mouse PCa cells which do not express UTY were knocked down for Utx (siUtx) and control (siNC) by siRNAs to assess changes in the abilities of proliferation and migration.

Results: RM-2 siUtx cells showed increased proliferation and migration compared with RM-2 siNC cells. By performing high-throughput RNA sequencing (RNA seq) and gene set enrichment analysis (GSEA) using mouse prostate tissues, we found that loss of UTX in the prostate activated inflammatory pathways, whereas inactivated DNA damage repair pathways. Immunostaining prostate tissues with an antibody to a macrophage marker, F4/80, exhibited significant accumulation of macrophages in UtxΔ, UtyΔ, p53+/+ mice compared to Utx+, UtyΔ, p53+/+ mice. In addition, immunofluorescent staining of irradiated prostate tissues with an antibody against a DNA repair marker, γH2AX, revealed that the time required for DNA damage repair was markedly prolonged in UtxΔ, UtyΔ, p53+/+ mice compared with Utx+, UtyΔ, p53+/+ mice.

Conclusions: These results suggest that loss of UTX contributes to prostate cancer development through promoting inflammation and impairing DNA damage repair, which may serve as novel therapeutic targets in PCa patients with UTX mutations.

Citation Format: NOBUHITO MURAMOTO, Masayuki Iwasaki, Yasuyuki Sera, Hisamitsu Ide, Shoichiro Mukai, Tsuyoshi Fukushima, Shigeo Horie, Hiroaki Honda. Elucidation of the pathogenesis of prostate cancer harboring mutations of epigenetic factor UTX using genetically engineered mice [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 1439.
Abstract 1442: Establishing a papillary renal cell carcinoma mouse model with activating Met mutation
Authors: Lal Preet , Ding Li , Chen Feng
Published: March 2024

Renal cell carcinoma (RCC) is a heterogeneous group of cancers of the nephron. Papillary renal cell carcinoma (PRCC) is the second most common type of RCC. c-Met activating mutations have been associated with Type 1 PRCC which are made up of small cells containing basophilic cytoplasm covering papillae and tubular structures. In patients with PRCC, c-MetH1112Y missense mutations are associated with both somatic and germline mutations. Previous functional studies indicate codon H1112 plays an important structural role in maintaining c-Met in the inactive conformation and alterations at this codon, including H1112Y, likely disrupt this interaction facilitating constitutive activation with increased c-MET activity. A c-MetH1112Y (c-MetHY) transgene with tetO promoter sequence was micro-injected into the mouse oocyte. Transgenic mice carrying this transgene have the ability to express this activated Met protein in cells expressing rtTA (reverse tetracycline-controlled transactivator) and when doxycycline is present. Mutant mice harboring this transgene along with additional trp53 mutations showed glomeruloid like Type 1 PRCC. Our in vivo studies confirmed the expression of c-MetHY gene within the kidney and Lrp2 and Ki67 staining indicated that PRCC arises from PT (proximal tubule). PRCC canonical markers CK7, AMACR and MET expression confirmed the Type1 PRCC in our mouse model. Additionally Pan-CK, E-Cad positive expression in mutant mice further confirmed PRCC arises from epithelial cells. We have generated and described type 1 PRCC mouse model with activating-MetHY mutation. This mouse model will help illustrate the oncogenic mechanism of Met activation and identify the critical events during progression of PRCC.

Citation Format: Preet Lal, Li Ding, Feng Chen. Establishing a papillary renal cell carcinoma mouse model with activating Met mutation [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 1442.
Abstract 145: Unraveling SMARCB1 depletion in hiPSC-derived neural crest cells: Implications for ATRT cell of origin
Authors: Wang Clark , Furnari Frank
Published: March 2024

Atypical teratoid rhabdoid tumors (ATRTs) are highly aggressive pediatric brain cancers that lack standardized treatment regimens and are associated with poor prognoses. Despite being driven by biallelic inactivation of a single gene, SMARCB1, ATRTs exhibit three epigenetic subgroups (SHH, TYR, and MYC), each with different clinical outcomes. Like many pediatric brain tumors, ATRTs share the fundamental underlying mechanism of stalled differentiation, suggesting that it may arise from neural progenitor cells that fail to terminally differentiate. However, unlike other brain tumors, ATRTs share molecular similarities with malignancies found outside of the CNS, especially in the kidney and soft tissues. This unique feature implies a putative cell of origin that is not restricted to the CNS: neural crest cells, a transient embryonic cell population that emerges from the neuroectoderm but then migrates throughout the embryo during development. To study SMARCB1 loss in a genetically defined neural progenitor cellular context with isogenic controls, the Furnari lab engineered a doxycycline inducible SMARCB1 knockdown system into human induced pluripotent stem cells (hiPSCs). After simultaneous SMARCB1 knockdown with directed neural differentiation, the resulting SMARCB1-depleted neural progenitor cells exhibited transcriptomic profiles that were similar to ATRTs, particularly to the SHH subgroup. Building upon these findings, the engineered hiPSCs were differentiated into neural crest cells using a modified version of a published protocol. Via RT-qPCR, these hiPSC-derived neural crest cells were confirmed to express typical neural crest marker genes and lose expression of SMARCB1 in the presence of doxycycline. Remarkably, ATP assays demonstrated that neural crest cells that were differentiated without SMARCB1 acquired enhanced proliferation in comparison to those differentiated with SMARCB1 intact. Further work aims to tease apart this proliferative phenotype in the neural crest cellular context via RNAseq, evaluate the impact of cell identity upon subgroup status, and to determine whether SMARCB1 depletion post-differentiation also enhances proliferation. Together, these findings feature the utility of hiPSC-derived models for investigating mechanisms underlying ATRTs to hopefully identify novel subgroup-specific targets.

Citation Format: Clark Wang, Frank Furnari. Unraveling SMARCB1 depletion in hiPSC-derived neural crest cells: Implications for ATRT cell of origin [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 145.