bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 5234: TGFbeta-conditioned K-NK cells are able to counter immune suppression in the tumor micro-environment
Authors: Kunert Andre , Oja Anna , Cesonyte Ieva , Franciszkiewicz Katarzyna , Sidhu Sukhvinder , Jetani Hardik , Bocconcelli Giulio , Igarashi Robert , Fantin Valeria R. 
Published: March 2024

Abstract
Recent advances have enabled the use of autologous K-NK cells to treat patients suffering from hematological malignancies. When aiming to treat patients suffering from solid tumors however, the suppressive effects of the tumor microenvironment (TME) and the involvement of other immune cells need to be considered to make a cell therapeutic potentially more effective.
TGFβ-conditioning (TGFβ-c), the co-culture of K-NK cells with low-dose TGFβ while undergoing cytokine-mediated expansion, imbues these cells with unique qualities suitable to mitigate immuno-suppressive effects of the TME, making them more efficacious toward solid tumors. The exposure to TGFβ during expansion results in significant downregulation of SMAD3, rendering these cells insensitive to TGFβ-mediated down-regulation of NK cell receptors. TGFβ-c K-NK cells display enhanced in vitro cytotoxicity towards 2D, 3D cultured cell lines as well as against organoid models derived of various indications. These cells infiltrate into patient-derived tumor organoids at higher frequency and have increased IFNγ secretion upon target-specific activation. The effect of the higher IFNγ output on a complementary anti-tumor immune subset was investigated in complex immune cell cultures and was indeed shown to enhance DC maturation.
In vivo studies revealed prolonged persistence of TGFβ-c K-NK cells in mice while retaining both altered gene-expression and functional hallmarks for up to 28 days, indicating that the conditioning results in a stable phenotype.
In summary, TGFβ-c K-NKs are a platform with enhanced potency to transition cell therapy from treating hematological to solid tumor malignancies.
Citation Format: Andre Kunert, Anna Oja, Ieva Cesonyte, Katarzyna Franciszkiewicz, Sukhvinder Sidhu, Hardik Jetani, Giulio Bocconcelli, Robert Igarashi, Valeria R. Fantin. TGFbeta-conditioned K-NK cells are able to counter immune suppression in the tumor micro-environment [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 5234.
Abstract 5251: RNA primed SMAR-T cells against multiple driver mutations, all HLA's designed for first line therapy
Authors: Slanetz Alfred E. , Srivatsa Sriram , Muzaffar Farzona , Barry Walter , Ryan David , Metri Salim , Slanetz Michael 
Published: March 2024

Abstract
Success of immune checkpoint inhibitors (e.g., anti-PD1 antibodies) have revolutionized cancer immunotherapy by demonstrating that a patient’s own T-cells recognize and treat cancer. The efficacy of PD-1 blockade is driven by recruitment of new T-cells from blood rather than via activation of pre-existing tumor infiltrating lymphocytes (Yost K.E., et al. Nat. Med. 2019). However, anti-PD1 therapy is most effective in ~5% of malignancies i.e., cancers with high mutational burden. Hence, the challenge of addressing most lower mutational burden cancers (the ~95%) needs an alternate treatment strategy. We address this challenge by using RNA to prime and expand peripheral T-cells to cancer-specific mutations ex-vivo. Using our proprietary, patented, and robust manufacturing method, we can generate T-cell populations reactive to as low as 8 and as high as 40 cancer-specific mutant proteins. In-vitro, these T-cells have cancer mutation-specific cytotoxicity and do not kill the normal cells. Further, the T-cells express homing receptors enabling them to infiltrate tumors and express high levels of TNFα and IFNγ, which are associated with effective tumor cytotoxicity and pro-inflammatory modification of tumor microenvironment (TME). Additional characterization shows these cells to be predominantly CD4+ and CD8+ T-cells bearing central and effector memory phenotypic with negligible regulatory or exhausted T-cells. We believe our T-cells can be used for cellular therapy in conjunction with, or as an alternative to, immune checkpoint inhibitors to treat lower mutational burden cancers present in most patients.
Citation Format: Alfred E. Slanetz, Sriram Srivatsa, Farzona Muzaffar, Walter Barry, David Ryan, Salim Metri, Michael Slanetz. RNA primed SMAR-T cells against multiple driver mutations, all HLA's designed for first line 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 5251.
Abstract 5253: Epigenetic drugs partially restored anti-cholangiocarcinoma activity of dysfunctional CD3+CD56+ cells
Authors: Wongkajornsilp Adisak , Chusorn Porncheera , Phongkitkarun Kriengkrai , Htwe Khin Su Su , Duangsa-ard Sunisa , Kasetsinsombat Kanda , Sawatpiboon Nathawadee 
Published: March 2024

Abstract
The cholangiocarcinoma cell-exposed cytokine-induced killer (CIK) cells or their most efficacious CD3+CD56+ subset displayed weaken anti-tumor cytotoxicity upon subsequent exposure. RNAseq analysis of the resistant phenotype gave rise to a list of drugs that could potentially counter the activity of genes associated with resistance to the anti-tumor activity of CIK cells. The top-ranking drugs appeared to be epigenetic drugs. Sublethal concentration of these drugs were assayed for their synergistic activity with CIK cell or the CD3+CD56+ subset for cytolysis of resistant CCA cells in tissue culture model. These drugs, although in low concentration, contained anti-tumor activity. The tumor cytolysis was greater after the combination with either CIK cells or the CD3+CD56+ subset. An in vivo model of CCA-implanted mice exhibited the synergistic activity of subtherapeutic dose of these drugs and CD3+CD56+ subset in suppressing tumor growth. It is concluded that epigenetic drugs in subtherapeutic dose could prevent the dysfunction of CCA-exposed CIK cells.
Citation Format: Adisak Wongkajornsilp, Porncheera Chusorn, Kriengkrai Phongkitkarun, Khin Su Su Htwe, Sunisa Duangsa-ard, Kanda Kasetsinsombat, Nathawadee Sawatpiboon. Epigenetic drugs partially restored anti-cholangiocarcinoma activity of dysfunctional CD3+CD56+ cells [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 5253.
Abstract 5352: SOX9-dependent immune microenvironment remodeling drives KRAS-induced lung adenocarcinoma progression
Authors: Menon Dinoop Ravindran , Zhong Hua , Ganesan Shridar , Sabaawy Hatim , Pine Sharon 
Published: March 2024

Abstract
Interactions between tumor intrinsic factors and tumor microenvironment play a vital role in disease progression and treatment response. SOX9, a transcription factor crucial for tissue development and homeostasis, has been linked to the advancement of tumor growth. However, its specific role as a driver in lung adenocarcinoma (LUAD) is not well-defined. Additionally, the impact of SOX9 on the tumor microenvironment has not been previously explored. We here identify SOX9 transcription factor to play a critical role in defining tumor immune microenvironment and driving disease progression. We employed CRISPR/Cas9 and Cre-LoxP gene knockout methods within the Kras G12D-induced mouse model of lung adenocarcinoma (LUAD). Our objective was to investigate the mechanisms through which SOX9 contributes to the development and advancement of lung adenocarcinoma. To substantiate our findings, we conducted immune profiling, gene expression analyses, RT-qPCR, and immunohistochemistry assessments in the Kras G12D-driven murine LUAD. These results were subsequently validated by examining bulk and single-cell gene expression profiles, as well as immunohistochemistry, in human lung adenocarcinoma. Our studies indicated that SOX9 played a very important role by enhancing lung tumor development, burden, and progression, leading to lower overall survival. SOX9. SOX9 consistently facilitated the growth of organoids in vitro. However, the promotion of tumor growth by SOX9 was notably reduced in immunocompromised mice compared to syngeneic mice. SOX9 demonstrated a suppressive effect on immune cell infiltration, particularly affecting the functionality of tumor-associated CD8+ T cells, natural killer cells, and dendritic cells. CD8+ T cells in the SOX9 overexpressing tumors showed upregulation of exhaustion markers. SOX9-mediated tumor growth advantage could not be observed in the CD8 knockout mice model, suggesting a CD8+ T cell-dependent role. In addition, SOX9 significantly upregulated collagen-related gene expression, leading to a substantial increase in collagen fibers possibly affecting the migration of immune cells into the tumor microenvironment. Together, our studies indicate that SOX9 plays a very critical role in modulating tumor microenvironment by controlling immune cell infiltration and CD8+ T cell exhaustion driving Kras G12D- driven lung tumor progression.
Citation Format: Dinoop Ravindran Menon, Hua Zhong, Shridar Ganesan, Hatim Sabaawy, Sharon Pine. SOX9-dependent immune microenvironment remodeling drives KRAS-induced lung adenocarcinoma progression [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 5352.
Abstract 5362: Targeting IRE1α alleviates the immunosuppressive tumor microenvironment in prostate cancer
Authors: Unal Bilal , Kuzu Omer Faruk , Jin Yang , Hurtado Daniel Camilo , Kuijjer Marieke Lydia , Daugaard Mads , Oo Htoo Zarni , Patterson John B. , Saatcioglu Fahri 
Published: March 2024

Abstract
Unfolded protein response (UPR) is a central stress response pathway in normal cells that is hijacked by tumor cells for their survival. However, how activation of UPR in cancer cells shapes the tumor microenvironment (TME) remain largely unexplored. Here, we investigated the role of IRE1α-XBP1s pathway on modulation of TME dynamics in prostate cancer (PCa). We found that IRE1α is increased in PCa patients and genetic or pharmacological inhibition of IRE1α signaling in the immunocompetent syngeneic mouse PCa model dramatically reduced tumor growth. Multiomics analysis of tumor samples upon IRE1α deletion in cancer cells showed significantly potentiated interferon (IFN) response and activation of immune system related pathways in the TME. Single-cell RNA sequencing (scRNA-seq) analysis revealed that the abundance of immunosuppressive cells, such as tumor-associated macrophages (TAMs) and T regulatory cells were markedly reduced in the IRE1α deficient tumors. Analysis of differentially expressed genes in the annotated cell types in the TME demonstrated that expression signatures associated with IFN responses were significantly enriched in TAMs, cancer cells, and dendritic cells. In addition, a novel scRNA-seq derived TAM gene signature is strongly associated with poor PCa survival. Furthermore, IRE1α inhibition by the small molecule MKC8866 (ORIN1001) that is in clinical trials significantly enhanced anti-PD-1 checkpoint inhibitor therapy in mice. Our findings indicate that IRE1α not only promotes cancer cell growth and survival, but it also strongly inhibits anti-tumor immunity in the PCa TME.
Citation Format: Bilal Unal, Omer Faruk Kuzu, Yang Jin, Daniel Camilo Hurtado, Marieke Lydia Kuijjer, Mads Daugaard, Htoo Zarni Oo, John B. Patterson, Fahri Saatcioglu. Targeting IRE1α alleviates the immunosuppressive tumor microenvironment in prostate 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 5362.
Abstract 5367: Single cell resolution spatial transcriptomics reveals genetic driver and TME interaction in clear cell renal cell carcinoma
Authors: Strunilin Ilya , Caravan Wagma , Abedin-Do Atieh , Houston Andrew , Li Yize , Song Yizhe , Cao Song , Mo Chia-Kuei , Chen Siqi , Pachynski Russel , Chen Feng , Ding Li 
Published: March 2024

Abstract
BAP1 mutations are associated with increased T cell and tumor-associated macrophage infiltration in clear cell renal cell carcinoma (ccRCC) and are predictive of poor prognosis. BAP1 mutations have also been found to be associated with T-cell infiltration in peritoneal mesothelioma and uveal melanoma. Because BAP1 mutations are known to be associated with aggressive disease and poor survival, we hypothesize that these mutations lead to a more immunosuppressive microenvironment. Using single nucleus RNA sequencing and high-resolution spatial transcriptomics, we confirmed that BAP1 mutant tumors are immune hot with a high fraction of Treg, exhausted CD8+ T cells, CD4+ T cells, and macrophages. At the same time, these tumors have a lower proportion of endothelial cells and pericytes. Evident of pro-tumor microenvironment we have found BAP1 mutations to be associated with the upregulation of SPP1, CD163, and MARCO immunosuppressive genes in tumor-associated macrophages. Here we propose a mechanism by which BAP1-mutant cancer cells induce an immunosuppressive microenvironment: through activation of the complement system and TGF-β signaling to attract macrophages. Taken together, our results suggest that BAP1 mutations contribute to poor ccRCC outcomes not only through cell-intrinsic effects, but also by promoting infiltration of Tregs and tumor-associated macrophages and polarizing infiltrated macrophages towards immunosuppressive phenotype.
Citation Format: Ilya Strunilin, Wagma Caravan, Atieh Abedin-Do, Andrew Houston, Yize Li, Yizhe Song, Song Cao, Chia-Kuei Mo, Siqi Chen, Russel Pachynski, Feng Chen, Li Ding. Single cell resolution spatial transcriptomics reveals genetic driver and TME interaction in clear cell renal cell carcinoma [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 5367.
Abstract 5372: KITENIN leads immunosuppression in the glioblastoma microenvironment via CCL20
Authors: Ahn Eun-Jeong , Kim Nah Ihm , Kim Sung Sun , Lee Jae-Hyuk , Rhee Joon Haeng , Kim Kyung-Keun , Kim Hangun , Lee Kyung-Hwa , Moon Kyung-Sub 
Published: March 2024

Abstract
Background: Glioblastoma (GBM) is characterized by an immunosuppressive tumor microenvironment (TME), limiting the effectiveness of existing treatments. The author investigated the role of KAI1 COOH-terminal interacting tetraspanin (KITENIN) in mediating this immunosuppression, focusing on its effect on cytokine secretion and tumor-infiltrating lymphocyte (TIL) profiles in the TME.
Materials and Methods: Using cytokine array and Luminex multiplex assays, the author evaluated cytokine secretion in KITENIN-modulated GL261 cells. The author further isolated TILs from an orthotopic mouse model implanted with these cells and induced in vitro differentiation of myeloid cells. The effect of neutralizing antibody for target cytokine, was assessed in vivo, and clinical relevance was investigated using GBM patient data from our cohort and TCGA.
Results: Employing cytokine array and Luminex multiplex assays, we found increased level of CC chemokine ligand 20 (CCL20) within KITENIN-overexpressed (KIT-HA) GL261 cell supernatants. Immunohistochemical analyses using GBM samples confirmed that both KITENIN and CCL20 expressions co-directionally increased and this was associated with decreased survival by TCGA analysis. Myeloid-derived suppressor cells (MDSCs), macrophages, and CD4+Foxp3+ regulatory T cells (Tregs) were observed to be higher in brain tumor implanted with KIT-HA GL261. Furthermore, KIT-HA GL261 conditioned medium expanded CD45+CD11b+Gr1-Ly6c+ monocytic MDSCs (M-MDSCs), an effect that was abrogated by CCL20 downregulation. In vivo neutralization of CCL20 resulted in reduced tumor volume, prolonged survival, and decreased M-MDSCs, thus affirming the role of CCL20 in mediating immunosuppression.
Conclusion: These findings underscore the KITENIN-CCL20 axis as a promising target for alleviating the immunosuppressive TME in GBM, potentially unlocking new avenues for GBM immunotherapy.
Citation Format: Eun-Jeong Ahn, Nah Ihm Kim, Sung Sun Kim, Jae-Hyuk Lee, Joon Haeng Rhee, Kyung-Keun Kim, Hangun Kim, Kyung-Hwa Lee, Kyung-Sub Moon. KITENIN leads immunosuppression in the glioblastoma microenvironment via CCL20 [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 5372.
Abstract 539: LN18-MRTX1719R cell line, a useful cell model for the discovery of the new PRMT5 inhibitors
Authors: Tang Yao , Wang Liping , Wang Guoqian , Ning Jinying , Hao Feng 
Published: March 2024

Abstract
In recent years, the PRMT5-MTA complex has emerged as a "synthetic lethal" novel target for the treatment of MTAP (methylthioadenosine phosphorylase)-deficient cancers. Kryukov et al. first reported the "synthetic lethal" effect of inhibiting PRMT5 in MTAP-deficient tumors, and they identified MTAP as the synthetic lethal partner of PRMT5 in 2016. MTAP is a tumor suppressor gene frequently lost in various cancers, including pancreatic cancer, non-small cell lung cancer, and mesothelioma. Furthermore, the loss of MTAP leads to the accumulation of MTA in cancer cells, which competes with PRMT5's co-factor SAM for binding and forms the PRMT5-MTA complex. Cancer cells lacking MTAP show enhanced sensitivity to PRMT5 loss. Therefore, this complex has emerged as a potential therapeutic target, particularly for the treatment of MTAP-deficient cancers. MRTX1719 is a highly effective selective inhibitor of the PRMT5-MTA complex, capable of selectively inhibiting PRMT5 activity in MTAP-deficient cells. As the most promising candidate drug, MRTX1719 is undergoing Phase I/II clinical evaluation for patients with MTAP-deficient cancers. Drug resistance is an inevitable problem with small molecule inhibitors. Currently, the mechanism of resistance to MRTX1719 is not clear, as MTAP/CDKN2A loss is an early driver event in most cancers. To further explore the resistance mechanism of MRTX1719, our group used the MTAP-deficient LN-18 cell line and induced mutagenesis using the mutagen ENU. We then used a drug low-concentration gradient method to establish the LN-18-MRTX1719R-resistant cell line. Specifically, LN-18 cells were treated with increasing doses of MRTX1719 for 6 months, and mixed clones that tolerated MRTX1719 were selected. Subsequently, single clones were selected, and a stable MRTX1719-resistant clone was obtained, named LN-18-MRTX1719R. Cell proliferation inhibition assay showed that the IC50 of LN-18-MRTX1719R was over 1000nM, which is 100-fold more resistant than the parental LN-18 cells (9.87 nM). We explored the resistance mechanisms using RNA-Seq and WES methods. The resistance to MRTX1719 of the LN-18-MRTX1719R cell line was highly stable after 20 cell passages. The LN-18-MRTX1719R cell line can be a useful cell line mode in the development of future PRMT5·MTA inhibitors.
Citation Format: Yao Tang, Liping Wang, Guoqian Wang, Jinying Ning, Feng Hao. LN18-MRTX1719R cell line, a useful cell model for the discovery of the new PRMT5 inhibitors [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 539.
Abstract 54: Engineering chimeric antigen receptor (CAR) T cells using autoantigen binding domains for treatment of B cell malignancies with increased specificity
Authors: Cheever Abiagil , Lindsay Hunter , Hansen MacKenzie , Kang Chloe , Weber K. Scott , O'Neill Kim 
Published: March 2024

Abstract
Introduction: Chronic inflammation often leads to cancer, and many autoimmune diseases lead to a significantly increased risk of developing cancer. Though many autoimmune diseases, like Graves’ Disease (GD), don’t have treatments that address this risk. Chimeric antigen receptor (CAR) T cells are an effective treatment for hematological cancers by eliminating all B cells, but a more targeted form of CAR T cells could eliminate specific B cell populations in B cell malignancies, including GD.
Methods: To allow our CAR T cells to target specific B cell populations that cause GD, we engineered chimeric autoantigen receptors (CAAR) with epitopes of thyroid stimulating hormone receptor (TSHR) as the binding domain. This acts as bait for the anti-TSHR B cells. Jurkat T cells were lentivirally transduced with these CAAR plasmids. Anti-TSHR B cell lines were engineered by transducing Nalm6 cells with lentivirus for anti-TSHR B cell receptors. Cytotoxicity assays were performed on primary human TSHR CAAR T cells incubated with the anti-TSHR B cell lines. Cytokine production, proliferation, and cytotoxicity assays were performed on these coincubated cells to determine the efficacy of our TSHR CAAR T cells.
Results: Two of the three Jurkat TSHR CAAR T cells showed significant binding to anti-TSHR antibodies (TRAbs). They also showed significant activation, measured by CD69 expression, when incubated with plate bound TRAbs, but not soluble TRAbs. The B cell lines were validated by binding to TSHR. The primary TSHR CAAR T cells showed significantly increased proliferation and production of TNF, IFN-y, and IL-2, when compared with controls. The TSHR CAAR T cells also had significant cytotoxic action against the anti-TSHR B cell lines, but not Nalm6 B cells. The cytotoxic function was comparable to our CD19 CAR T cell with the same activation domains, though it demonstrated specificity to only anti-TSHR B cells.
Conclusion: TSHR CAAR T cells are a potentially curative treatment for GD that will stop chronic inflammation and could reduce GD patient’s risk of developing thyroid cancer. This is also an important development in CAR T cell therapies that allows for specific targeting of certain B cell populations.
Citation Format: Abiagil Cheever, Hunter Lindsay, MacKenzie Hansen, Chloe Kang, K. Scott Weber, Kim O'Neill. Engineering chimeric antigen receptor (CAR) T cells using autoantigen binding domains for treatment of B cell malignancies with increased specificity [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 54.