bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2064: Targeting Cblb-Notch1 axis as a novel strategy for cancer immunotherapy
Authors: Monticone Giulia , Csibi Fred , Leit Silvana , Austin Jermaine E. , Ucar Deniz A. , Hossain Fokhrul M. , Majumder Samarpan , Osborne Barbara A. , Loh Christine , Miele Lucio 
Published: June 2022

Abstract
Background: Tumor-induced immunosuppression is a way in which cancers evade the host immune response. In recent years, a great effort has been made in designing immunotherapies that can boost the immune system response against cancer. However, not all tumors respond to these therapies. Therefore, there remains an unmet need for immunotherapies that are able to circumvent tumor-induced immunosuppression. Adenosine is an immunosuppressive metabolite which is overproduced in hypoxic tumor microenvironments and dampens T-cell anti-tumor immune responses. Adenosine A2A receptor (A2AR) activation was shown to downregulate Notch1, a key regulator of T-cell functions, in CD8+ T-cells, leading to immunosuppression. Notch1 signaling appears to protect T-cells against immunosuppressive signals, as Notch1 overexpressing T-cells were shown to be resistant to tumor-induced immunosuppression mediated by myeloid-derived suppressor cells (MDSC), as well as adenosine. We hypothesize that rescuing Notch1 from downregulation will make T-cells resistant to tumor-induced immunosuppression. We applied several functional assays in primary CD8+ T-cells and tumor-derived organoids to study how Notch1 is regulated by A2AR and explore strategies to target this pathway for cancer immunotherapy.
Results: Our results indicate that the ubiquitin ligase Cbl-b, a negative regulator of T-cell functions, is responsible for the ubiquitination and degradation of Notch1 in CD8+ T-cells. A2AR signaling leads to Notch1 downregulation by promoting Cbl-b-mediated Notch1 degradation. Inhibition of Cbl-b using small molecule compounds restores Notch1 and T-cell functions in CD8+ T-cells in the presence of adenosine, making them resistant to A2AR-mediated immunosuppression. Cbl-b inhibitors show anti-tumor activity in tumor-derived organoids from pre-clinical models and enhance immune-checkpoint immunotherapy, by promoting T-cells anti-cancer responses.
Conclusions: Our work suggests that promoting Notch1 signaling by blocking Cbl-b-mediated degradation results in increased T-cell responses and resistance to immunosuppression. Targeting Cbl-b-Notch1 axis represents a promising novel strategy to boost anti-cancer T-cell responses.
Citation Format: Giulia Monticone, Fred Csibi, Silvana Leit, Jermaine E. Austin, Deniz A. Ucar, Fokhrul M. Hossain, Samarpan Majumder, Barbara A. Osborne, Christine Loh, Lucio Miele. Targeting Cblb-Notch1 axis as a novel strategy for cancer immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2064.
Abstract 209: Elucidating the role of NMDA subunit NR2B in non-tumor and tumor-bearing mice
Authors: Castro Jesus , Gaelzer Mariana , Welford Scott 
Published: June 2022

Abstract
Therapeutic options for glioblastoma multiforme (GBM), the most common and fatal brain tumor, have been limited over the course of recent decades, leaving radiation as one of the few effective therapies. While advancements in radiation therapy have improved life expectancy and patient outcome, exposure to radiotherapy causes normal tissue toxicity and damage to the central nervous system, resulting in cognitive impairment and negative patient side effects. In the hippocampus, the memory processing center of the brain, radiotherapy-induced elevated glutamate levels lead to over-excitation of N-methyl-D-aspartate receptors (NMDAR) and thus cell death. To investigate the NMDAR, we repurposed a NMDA subtype specific inhibitor (Ifenprodil) and treated non-tumor mice before radiation. We found that Ifenprodil rescues cell death and cognition by inhibiting the NR2B subunit of NMDA receptors and preventing over-excitement-induced excitotoxicity, despite presence of high glutamate levels. In order to relieve glioma patients of the negative side effects of radiotherapies, we aim to further investigate the critical role of NR2B inhibition in non-tumor and tumor-bearing mice to reduce excitotoxicity and prevent damage to the central nervous system, opening avenues for therapeutic targeting and drug development. To accomplish this, we designed promotor-specific plasmids, only activated when present in their designed neuronal cell types, to initiate CRISPR/Cas9 genome editing and knock out the NR2B subunit of NMDAR. With NR2B knocked out across 5 different cell types, we exposed hippocampal organotypic slice cultures, derived from non-tumor mice, to 2 Gy radiation and ran flow cytometry to reveal which neuronal NR2B subunits play critical roles in excitotoxic phenotypes. To further these findings, we utilized a recently developed in-utero-electroporation mouse model that produces glioblastomas in immunocompetent mice by inactivating select tumor-suppressor genes via CRISPR/Cas9 genome editing at embryonic day 14.5. By applying different CRISPR/Cas9 gRNAs into our tumor-bearing mice, we can not only explore the robust role of NR2B inhibition in tumor brains with varying expressions, but also shed light onto how NR2B inhibition protects normal brain while leaving tumors radiosensitive. In conclusion, this work seeks to further develop our understanding of radiotherapy-induced cytotoxicity in order to prevent cognitive impairment and negative side effects in patients, improving their quality of life. Ultimately, the results from our findings will lead to the advancement of targeted drug development and therapeutic options.
Citation Format: Jesus Castro, Mariana Gaelzer, Scott Welford. Elucidating the role of NMDA subunit NR2B in non-tumor and tumor-bearing mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 209.
Abstract 2090: Humanized IL36R mice provide a preclinical tool for the evaluation of therapeutic drugs targeting IL36R
Authors: Zhou Xiaofei , Chang Jiahui , Frame Jenna , Li Chong , Bai Zhifang , Shen Yuelei 
Published: June 2022

Abstract
The IL36 family of cytokines (IL36a, IL36b, and IL36g) is part of the IL-1 superfamily and triggers proinflammatory signaling through the IL36 receptor complex. This heterodimer complex is formed by the IL36 Receptor (IL1RL2) and IL-1R co-receptor accessory protein (IL-1RAcP). IL36R is present on a diverse variety of cell types, including the surface of dendritic cells, CD4+ T cells, and epithelial cells in various tissues. IL36 signaling can exert widespread inflammation, and its activity is implicated in several inflammatory diseases affecting epithelial tissues, including inflammatory bowel disease, rheumatoid arthritis, and psoriasis. Thus, identification of novel therapeutics targeting this pathway are of significant clinical interest. To provide a preclinical platform for efficacy assessments of novel anti-human IL36R drugs, we generated a humanized hIL36R knock-in mouse model. In this model, a chimeric IL1RL2 coding sequence, which is composed of the human IL1RL2 extracellular region and the mouse Il1rl2 cytoplasmic region, was inserted after the 5’UTR of mouse Il1rl2 gene using CRISPR-Cas9-based gene editing technology. The mouse cytoplasmic region was maintained to ensure intact downstream signaling. Polymerase chain reaction assessment of mRNA isolated from the lung tissue of edited homozygous humanized IL36R mice confirmed the absence of mouse Il1rl2 transcripts. Finally, to confirm the expression of human IL36R protein, immunohistochemistry was performed on the small intestine and skin tissue of humanized mice subjected to a chemical induction of psoriasis. Psoriasis-like symptoms were achieved by application of imiquimod cream on the back skin of the mice for 5 consecutive days. As expected, IL36R protein expression was detected in both tissues isolated from the homozygous humanized mice. These results indicate the successful generation of humanized IL36R mice, and suggest that this model will be an effective tool for future in vivo efficacy evaluation of immunomodulatory drugs targeting human IL36R.
Citation Format: Xiaofei Zhou, Jiahui Chang, Jenna Frame, Chong Li, Zhifang Bai, Yuelei Shen. Humanized IL36R mice provide a preclinical tool for the evaluation of therapeutic drugs targeting IL36R [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2090.
Abstract 2098: Practical cell counting method selection to increase the quality of cell counting results
Authors: Chan Leo , Bell Jordan , Lin Bo 
Published: June 2022

Abstract
The importance of cell counting has increased significantly in the last decade due to the major advances in the fields of cell and gene therapy, biologics production, and regenerative medicine. This has necessitated the development of a standardized approach to cell counting assays. In recent years, the U.S. Food and Drug Administration (FDA), in collaboration with the National Institute of Standards and Technology (NIST) and the International Organization for Standardization (ISO), has launched an effort to standardize cell counting methods to improve the confidence in cell counting measurements. There is a wide range of biological sample types, various formulations, and bioprocessing steps for cell and gene therapy products. Furthermore, there are no ground truth reference materials for live cells, making the determination of accuracy of cell counting difficult defined in the ICH Q2 (R1). Therefore, in order to increase the confidence of cell counting results, it is critical to follow the two recently published ISO cell counting standards to ensure cell counting methods are tailored to the specific sample and purpose.In this work, we will provide insights and guidance for the ISO standards for method selection. The important aspects of the ISO cell counting standards have been distilled to six key factors for selecting the cell counting methods. Attention to these details will allow researchers to more easily adhere to the ISO recommendations to ensure high quality cell counting measurements, and in doing so, sites can meticulously document their rationale, process, and data results.
Citation Format: Leo Chan, Jordan Bell, Bo Lin. Practical cell counting method selection to increase the quality of cell counting results [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2098.
Abstract 2192: HSF1 downregulation of CCL5 reduces CD8 + T cell trafficking in breast cancer
Authors: Jacobs Curteisha LeeAnn , Shan Sakhi , Lu Wen-Cheng , Carpenter Richard 
Published: June 2022

Abstract
Breast cancer is the second leading cause of cancer-related death in women. The presence of cytotoxic immune cells, specifically CD8+ T cells, in breast tumors is associated with better patient outcomes. Understanding the mechanisms that regulate the infiltration of CD8+ T cells into breast tumors could improve the treatment and enhance patient overall survival. Utilizing a novel HSF1 activity gene signature, we observed that HSF1 activity was negatively associated with the presence of CD8+ T cells in breast cancer patients. Both CIBERSORT analysis of TCGA data as well as direct assessment of patient primary specimens demonstrated that patient tumors with high HSF1 activity had lower numbers of CD8+ T cells. To functionally test this relationship, HSF1 was knocked down in 4T1 breast cancer cells and grown orthotopically in BALB/c mice. Tumors with HSF1 knockdown showed lower tumor volumes and increased CD8+ T cell infiltration. We further tested the effect of CD8+ T cells on the growth of HSF1 knockdown tumors by depleting CD8+ T cells in BALB/c mice. HSF1 knockdown tumors were significantly larger with CD8+ T cell depletion suggesting a functional role for HSF1 to inhibit CD8+ T cell infiltration and protect the tumor from immune-mediated killing. To investigate the mechanism by which HSF1 affects CD8+ T cells, we investigated whether HSF1 affects chemotactic cytokines. We found that loss of HSF1 significantly increased secretion of CCL5, a known chemoattractant for CD8+ T cells. Loss of HSF1 also increased mRNA levels of CCL5, suggesting a transcriptional effect on CCL5. To test the importance of CCL5 in the phenotype of HSF1 knockdown, a transwell migration assay was performed wherein T cells from BALB/c mice were placed in the upper chamber of a transwell chamber and the lower chamber contained conditioned medium from 4T1 cells with knockdown of HSF1 with or without knockdown of CCL5. The lower chamber was subjected to flow cytometry for CD3/CD8 to identify CD8+ T cells and we observed that loss of HSF1 increased migration of CD8+ T cells and this was ablated with additional loss of CCL5. These results suggest CCL5 is a significant mediator for the recruitment of CD8+ T cells when HSF1 is lost in cancer cells. We propose the model whereby high HSF1 activity in breast cancer cells suppresses expression and secretion of the chemo-attractant cytokine CCL5 that ultimately leads to a decrease in CD8+ T cells in breast tumors microenvironment.
Citation Format: Curteisha LeeAnn Jacobs, Sakhi Shan, Wen-Cheng Lu, Richard Carpenter. HSF1 downregulation of CCL5 reduces CD8 + T cell trafficking in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2192.
Abstract 2200: Multilineage plasticity in prostate cancer through expansion of stem-like luminal epithelial cells with elevated inflammatory signaling
Authors: Zaidi Samir , Zhao Jimmy , Chan Joseph , Martine Roudier , Wadosky Kristine , Gopalan Anuradha , Karthaus Wouter , Watson Philip , True Lawrence , Nelson Peter , Scher Howard , Morris Michael , Haffner Michael , Goodrich David , Pe'er Dana , Sawyers Charles 
Published: June 2022

Abstract
Lineage plasticity is a well-established mechanism of resistance to targeted therapies in lung and prostate cancer, where tumors transition from adenocarcinoma to small-cell or neuroendocrine carcinoma. Single-cell analysis of a cohort of late stage castration-resistant human prostate cancers (CRPC) revealed a greater degree of plasticity than previously appreciated, with multiple distinct neuroendocrine (NEPC), mesenchymal (EMT-like), and other subpopulations detected within single biopsies. To explore the steps responsible for initiation of this process, we utilized two genetically engineered mouse models of prostate cancer that recapitulate progression from adenocarcinoma to neuroendocrine disease. Time course studies reveal expansion of stem-like luminal epithelial cells (Sca1+, Psca+, called L2) that, based on trajectories, gave rise to at least 4 distinct subpopulations, NEPC (Ascl1+), POU2F3 (Pou2f3+), TFF3 (Tff3+) and EMT-like (Vim+, Ncam1+). Such populations are also seen in human prostate and small cell lung cancers. Furthermore, transformed L2-like cells express stem-like and gastrointestinal endoderm-like transcriptional programs, indicative of reemerging developmental plasticity programs, as well as elevated Jak/Stat, interferon, and FGF pathways. Strikingly pharmacologic inhibition of Jak/Stat and FGFR results in reversal of plasticity states and subsequent sensitivity to androgen receptor inhibitors (ARSIs). In sum, while the magnitude of multilineage heterogeneity, both within and across patients, raises considerable treatment challenges, the identification of highly plastic luminal cells as the likely source of this heterogeneity provides a target for more focused therapeutic intervention.
Citation Format: Samir Zaidi, Jimmy Zhao, Joseph Chan, Roudier Martine, Kristine Wadosky, Anuradha Gopalan, Wouter Karthaus, Philip Watson, Lawrence True, Peter Nelson, Howard Scher, Michael Morris, Michael Haffner, David Goodrich, Dana Pe'er, Charles Sawyers. Multilineage plasticity in prostate cancer through expansion of stem-like luminal epithelial cells with elevated inflammatory signaling [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2200.
Abstract 2212: Racially disparate serum levels of inflammatory cytokines, satiety and stress hormones, and exosomal microRNAs in women with or without a breast cancer diagnosis
Authors: Sudan Sarabjeet Kour , Vikramdeo Kunwar Somesh , Sharma Amod , Deshmukh Sachin Kumar , Srivastava Sanjeev Kumar , Poosarla Teja , Holliday Nicolette P. , Dyess Donna L. , Singh Ajay P. , Singh Seema 
Published: June 2022

Abstract
Breast cancer (BC) is the most common non-cutaneous malignancy and the second leading cause of cancer-related death in American women. Health disparities in incidence and clinical outcome are also reported among different racial groups, most notably African American (AA) women, who are often diagnosed at a young age with aggressive BC and exhibit greater mortality than Caucasian American (CA) women. Since socioeconomic difficulties can have tremendous impact on psychophysiology besides limiting the access to optimal healthcare, we examined the serum levels of stress (cortisol) and satiety (leptin) hormones as well as inflammatory cytokines (resistin and interleukin-6/IL-6) in AA and CA women. To observe a potential epigenomic connection, we also performed profiling of a targeted set of exosomal microRNAs in serum samples. The study was conducted under an Institutional Review Board (IRB) approved protocol. All subjects participated voluntarily, and their consents were obtained. Serum levels of resistin, IL-6, leptin, and cortisol were quantified by Enzyme linked Immunosorbent Assay (ELISA) using commercial kits. Exosomes were isolated using precipitation method and recovered by standard centrifugation. Total RNA was isolated from exosomal preps and subjected to stem-loop RT-PCR for quantitation of a set of inflammation-associated microRNAs. We found that the levels of resistin, leptin, IL-6 and cortisol were higher in women with a BC diagnosis than non-BC subjects. Moreover, AA women with or without BC showed significantly higher levels of these cytokines and hormones in their serum as compared to the CA women with or without a BC diagnosis, respectively. We also observed differential expression of several microRNAs in serum of BC women as compared to their normal counterparts, of which five (miR511, miR33a, miR27a, miR6794, miR143-3p) exhibited highest presence in serum exosomes of AA women with BC. Together, these findings suggest that relatively greater exposure of minority women to social stressors may have epigenomic consequences and may potentially be linked to the observed BC racial health disparities.
Citation Format: Sarabjeet Kour Sudan, Kunwar Somesh Vikramdeo, Amod Sharma, Sachin Kumar Deshmukh, Sanjeev Kumar Srivastava, Teja Poosarla, Nicolette P. Holliday, Donna L. Dyess, Ajay P. Singh, Seema Singh. Racially disparate serum levels of inflammatory cytokines, satiety and stress hormones, and exosomal microRNAs in women with or without a breast cancer diagnosis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2212.
Abstract 2221: Molecular analysis finds excision margin width predictive of recurrence in acral melanoma
Authors: Attrill Grace Heloise , Bailey Teresa A. , Lum Trina , Conway Jordan W. , Long Georgina V. , Wilmott James S. , Scolyer Richard A. 
Published: June 2022

Abstract
Surgical excision with histologically clear margins results in cure of most melanoma patients. Involvement of the margins is associated with tumour recurrence and adverse outcomes. Determining whether or not the histological margins are involved by melanoma, particularly in situ melanoma, can be very difficult particularly in acral locations where insitu melanoma can be extremely subtle. This study sought to determine the proximity of acral melanoma in situ to the surgical excision margin using multiple methods including routine histopathological assessment, FISH (CCND1 amplification), and multiplex immunofluorescence (mIF: SOX10, p16, Cyclin D1 and PRAME) in primary acral melanomas with initially histopathologically reported clear excision margins of &gt;0.3mm (n=85). The clear excision margins determined by each modality were compared between those melanomas which recurred (n=47) and those which remained recurrence-free at last follow-up (median = 46 months, range = 1-260 months) (n=35). Histopathologically confirmed excision margin width was smaller in the tumours of patients who recurred (mean = 7.083mm vs 9.964mm, p = 0.0811). Intratumoural CCND1 amplification was increased in recurrence-free patients in comparison to recurrence patients (16.2% vs 7.7%). FISH and mIF confirmed the correlation of intratumoural CCND1 amplification with Cyclin D1 expression SOX10+ melanoma cells (p=0.0347). No correlations between recurrence status and Cyclin D1, PRAME or p16 expression levels in the tumour or epidermis were identified. In histopathologically normal epidermal tissue as assessed on H&E sections,Cyclin D1+, p16- and PRAME+ cells were classified as malignant cells when co-expressed with SOX10. Upon measuring the distance from the tumour margin to the nearest malignant cell, we found that malignant cells were closer to the tumour in recurrence patients in comparison to recurrence-free patients (mean = 7.324mm vs 9.464mm, p = 0.2735). These distances closely correlated with the excision margin width as determined via histopathological assessment (R2 = 0.7424, p&lt;0.0001). In conclusion, histological assessment, FISH and mIF determined excision margins are highly correlative in acral melanomas. Narrower excision margins were associated with local recurrence of melanoma.
Citation Format: Grace Heloise Attrill, Teresa A. Bailey, Trina Lum, Jordan W. Conway, Georgina V. Long, James S. Wilmott, Richard A. Scolyer. Molecular analysis finds excision margin width predictive of recurrence in acral melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2221.