bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 1542: MALAT1 ablation dismantles homologous recombination repair machinery and sensitizes castrate resistant prostate cancer cells to PARP inhibitor
Authors: Yadav Anjali , Biswas Tanay , Praveen Ayush , Ateeq Bushra 
Published: June 2022

Abstract
Poly (ADP-ribose) polymerase (PARP) inhibitors have emerged as a promising target of intervention for metastatic castration-resistant prostate cancer (mCRPC), as approximately one-third of mCRPC patients harbor mutations in the genes associated with the homologous recombination (HR) pathway. Nonetheless, most patients inevitably develop resistance to PARP inhibition (PARPi), due to the induction of reversion mutations in the HR pathway. These mutations restore the function of HR genes and revert to the HR-proficient stage, which in turn decreases the vulnerability of cancer cells to PARP inhibitors. Here, we report that the clinical utility of PARPi could be efficaciously extended by targeting Metastasis associated lung adenocarcinoma transcript (MALAT1), a long non-coding RNA (lncRNA) often elevated in advanced-stage prostate cancer (PCa). In support of this, we show that mCRPC patients exhibit higher expression of genes involved in HR, which positively correlate with MALAT1 levels. Furthermore, RNA interference (RNAi)-mediated depletion of MALAT1 in CRPC cells perturb the expression of key HR genes, namely BRCA1/2, RAD51, EXO1, CHEK1/2, subsequently resulting in HR deficiency. This, in turn, escalates accumulation of DNA lesions in the MALAT1 ablated cells as noticed by abundance in γH2AX, a marker for DNA damage. As a consequence, MALAT1 ablated cells instigates G1/S phase arrest to fetch additional time to repair the damaged DNA or to induce apoptosis. Furthermore, we also provide evidence that the HR deficiency induced by MALAT1 depletion phenocopies “BRCAness” and exhibits synergy with clinically approved DNA repair inhibitors such as Olaparib. In particular, co-inhibition of MALAT1 and PARP1 exhibits a significant anti-proliferative effect reduced clonogenic survival and delays the resolution of γH2AX foci in CRPC cell lines. Taken together, our results establish that MALAT1 plays a central role in regulating the DNA damage response and provides a mechanistic rationale for dual targeting of MALAT1 and PARP in mCRPC patients.
Citation Format: Anjali Yadav, Tanay Biswas, Ayush Praveen, Bushra Ateeq. MALAT1 ablation dismantles homologous recombination repair machinery and sensitizes castrate resistant prostate cancer cells to PARP inhibitor [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 1542.
Abstract 1578: Interaction of E-cadherin and EGFR in the collective cell migration of colon cancer cells
Authors: Winarso Viriya Adhiguna , Lu Yi-Wen , Weng Yu-Cheng , Hou Xiang-Ling , Koo Hui Min , Li Yu-n , Chao Wei-Ting 
Published: June 2022

Abstract
Collective cell migration is one of the most important cellular events where a cluster of cells moves together in a coordinated way. Many studies have found this phenomenon as the hallmark of cancer progression and metastasis. In epithelial single cell migration, downregulation of adherens junction (AJ) molecule E-cadherin occurs at the initial step known as the Epithelial to Mesenchymal Transition (EMT). However, in collective cell migration, E-cadherin junction is maintained to provide adhesion between cells. Moreover, it also mediates the cell-cell intercommunication that is critical, for instance, to alter the group polarization through actin cytoskeleton remodeling. Interestingly, Epidermal Growth Factor Receptor (EGFR) was found to colocalize with E-cadherin in the cell-cell contact region, implicating that E-cadherin and EGFR might interact in the cell junction of collective cell migration. To address this, a human colorectal cancer cell line exhibiting epithelial phenotype HT-29 was used. The cell density and calcium presence were modified to infer the effect of differential expression and stability of E-cadherin on the cellular signaling of proliferation and migration. The results showed that stable and enhanced expression of E-cadherin downregulates EGFR expression and reduces its activity. Moreover, the Src phosphorylation increased when the cell-cell contact was strengthened via calcium addition and decreased when the cell-cell contact was diminished via calcium chelation. Overall, this indicates that E-cadherin expression at the cell-cell junction and contact stability determine the EGFR expression, activity, and Src phosphorylation. With the goal of deciphering the relationship of E-cadherin and EGFR in cell contact, proliferation, and migration, we intend to finally implement the knowledge as a strategy or approach in promoting alternative methods to inhibit metastasis.
Citation Format: Viriya Adhiguna Winarso, Yi-Wen Lu, Yu-Cheng Weng, Xiang-Ling Hou, Hui Min Koo, Yu-n Li, Wei-Ting Chao. Interaction of E-cadherin and EGFR in the collective cell migration of colon cancer cells [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 1578.
Abstract 1600: Dose-dependent effects of interferon-gamma treatment on pancreatic ductal adenocarcinoma cell states
Authors: Liberman Deborah A. , Barkley Dalia , Yanai Itai 
Published: June 2022

Abstract
Interferon-gamma (IFNγ) is a canonical proinflammatory cytokine that has well-studied antitumor functions such as the upregulation of MHC Class I (MHC-I), increased Th1 development, macrophage activity, innate immune cell recruitment, and in tumor cells increased apoptosis, decreased angiogenesis and proliferation. However, IFNγ is also important in limiting tissue destruction due to inflammation, taking on an immunotolerant role and allowing cancer cells to evade immune surveillance. We previously identified an interferon response cell state across diverse cancer types, as defined by the relative expression of IFIT1, HLA-A, HLA-DRA, STAT1, and IRF1. The interferon response cell state is heterogeneously expressed in cancer cells and appears to function both by cancer cell extrinsic and intrinsic mechanisms. Several clinical trials have tested the efficacy of IFNγ as a therapeutic for immune system stimulation in patients with melanoma. However, these have failed to detect effective outcomes, and have even noted some pro-growth actions, perhaps consistent with IFNγ’s immunotolerant role in maintaining tissue homeostasis. We are interested in identifying cell populations that are vulnerable to the antitumor functions of IFNγ, as well as cell populations that may be susceptible to IFNγ’s pro-tumor actions. To test this, we treated a pancreatic ductal adenocarcinoma cell line with three doses of IFNγ in vitro every day for five days and analyzed expression of proteins associated with IFNγ signaling by flow cytometry each day. Distinct doses of IFNγ have been shown to affect varying degrees of antitumor and pro-tumor responses. For example, growth inhibition has been seen upon treatment with high dose IFNγ, while treatment with low dose IFNγ is consistent with increased metastatic capability such as increased resistance to cytotoxic lymphocytes and natural killer cells despite the upregulation of MHC-I. We thus treated cells with three doses of IFNγ: low, normal, and high. Preliminary results have shown a dose dependent effect of IFNγ including the upregulation of MHC-I and PDL1. Ongoing work includes identifying changes in cell states using single-cell RNA-sequencing to uncover how cancer cells respond to varying intensities of IFNγ exposure. Additionally, we are analyzing how release of IFNγ from cancer cells changes following treatment of IFNγ at different doses. This work will provide insight into the mechanisms by which cancer cells evade immune surveillance and how cell states can describe a tumor’s vulnerability to IFNγ’s antitumor and pro-tumor effects.
Citation Format: Deborah A. Liberman, Dalia Barkley, Itai Yanai. Dose-dependent effects of interferon-gamma treatment on pancreatic ductal adenocarcinoma cell states [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 1600.
Abstract 1624: Immune cell killing assay for measuring the effects of immunomodulating agents in lung cancer cells in vitro
Authors: Mäki-Jouppila Jenni H. , Rissanen Jukka P. , Fagerlund Katja M. 
Published: June 2022

Abstract
Direct effects of cancer therapeutics on immune cell activity can be challenging to determine on system level in vivo. Therefore, it is important to develop and discover novel ways to study the effects of immune cells on cancer cells using also in vitro assays. Our aim was to validate an in vitro immune cell killing assay where activated peripheral blood mononuclear cells (PBMCs) target lung cancer cells and induce apoptosis.
A549 human lung carcinoma cells (ATCC) were used as target cells in the co-culture and were plated into 96-well plates. 24 hours later, human PBMCs (Lonza) were added to the culture as effector cells and activated with anti-CD3 and IL2. Apoptosis was detected using IncuCyte Annexin V reagent, which was added to the cells at the same time with immune cell activators. Cell proliferation was monitored with a live cell imager IncuCyte S3 (Sartorius) for three to four days before terminating the cultures. Cell viability was determined by CellTiter Glo (Promega) at the end of the study.
Treatment of cells with anti-CD3 was able to activate immune cells alone but the effects were observed with lower test concentrations in combination with IL-2. Therefore, IL-2 potentiates the effect of anti-CD3 on immune cell activation and decreases cancer cell viability in co-culture.
The immune cell killing assay based on live cell imaging can be used for studying the effects of immunomodulating agents on cancer cell viability and to evaluate the activity of immune cells in the presence of therapeutics.
Citation Format: Jenni H. Mäki-Jouppila, Jukka P. Rissanen, Katja M. Fagerlund. Immune cell killing assay for measuring the effects of immunomodulating agents in lung cancer cells in vitro [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 1624.
Abstract 1631: Anti-cancer effects of vorinostat on 3D cultured pancreatic cancer cells
Authors: Emamzaden Fatemeh Nouri , Word Beverly , Hammons George , Lyn-Cook Beverly D. 
Published: June 2022

Abstract
Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive cancers, often having a low survival rate mainly due to late diagnosis and limited therapies because of drug resistance. Treatment results shown in cell models used in studying various drugs often fail in clinical trials. The lack of translation of the effects observed in vitro to patients continues to halt progress. Genetic mutations are known in PDAC; however, recent studies have suggested the role of epigenetics in its initiation and progression to a malignant phenotype. Histone deacetylase (HDAC) proteins are highly expressed in a variety of cancers, including pancreatic cancer. Recent studies have shown that targeting these proteins through the use of HDAC inhibitors may be an effective therapy for pancreatic cancer. Using a 3D cultured pancreatic cell model, anti-cancer effects by vorinostat, a HDAC inhibitor, were investigated. Although both pancreatic cell lines, PANC-1 and Mia, contain mutated K-ras and P53, a differential effect in response to vorinostat was observed in expression of cancer genes using a Human Pancreatic Adenocarcinoma TagMan Array 96 (Applied Biosystems). In the PANC-1 cell model, vorinostat decreased expression of STAT6, which is known to play a role in migration and invasion. It also decreased significantly expression of insulin growth factor 1 (IGF1), IL-6, EGF, VEGFA and TGFB1. CCND2 and CCND2 were also downregulated. In contrast, Mia cell model showed decreased expression of EGF and IGF1, but no change with the other genes. These data suggest that vorinostat has effects on several potential pathways in pancreatic cancer possibly through epigenetic mechanisms that affect certain genes involved in progression and invasion. Currently, this drug in combination with other drugs is being investigated for pancreatic cancer.
Citation Format: Fatemeh Nouri Emamzaden, Beverly Word, George Hammons, Beverly D. Lyn-Cook. Anti-cancer effects of vorinostat on 3D cultured pancreatic cancer cells [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 1631.
Abstract 1642: Evaluating in vivo efficacy of anti-VSIG4 antibodies in humanized B-hVSIG4 mice
Authors: Lyu Ruili , Shang Chengzhang , Kokabee Leila , Nie Yanhui , Yang Yi 
Published: June 2022

Abstract
V-set and immunoglobulin domain containing 4 (VSIG4) protein is an immune checkpoint molecule that is related to the B7 family of immune regulatory proteins. B7 family members have been implicated in both positive and negative regulation of T cell activation, particularly VSIG4 is a known negative regulator of T cell proliferation and IL-2 production. Pathological consequences of VSIG4 expression are associated with inflammatory disorders, lung cancer development, high-grade glioma, and multiple myeloma, suggesting VSIG4 is a potential target for drug development. Therefore, Biocytogen developed humanized VSIG4 mice (B-hVSIG4 mice) by replacing the murine Vsig4 gene with the human VSIG4 gene through homologous recombination. Flow cytometry analysis showed human VSIG4 was detectable in peritoneal exudate macrophages (PEMs) in homozygous B-hVSIG4 mice compared to wild type mice. Additionally, percentages of T cells, B cells, NK cells, DCs, granulocytes, monocytes, macrophages, CD8+ T cells, CD4+ T cells and Tregs in homozygous B-hVSIG4 mice were similar to those in wild type mice, demonstrating that introduction of human VSIG4 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types. For in vivo efficacy evaluation of an anti-human VSIG4 antibody (Ab-A, synthesized in house) using B-hVSIG4 mice, mouse colon adenocarcinoma MC38 cells were subcutaneously implanted into homozygous B-hVSIG4 mice (female, 7-week-old, n=5). Mice were grouped and treated when the tumor volume reached approximately 100 mm3. Results showed that an anti-hVSIG4 antibody was efficient at inhibiting tumor growth in B-hVSIG4 mice. In conclusion, B-hVSIG4 mice are as a promising model for preclinical in vivo studies to assess pharmacodynamic of anti-hVSIG4 antibodies. Keywords: VSIG4, humanized mice, immune checkpoint molecule
Citation Format: Ruili Lyu, Chengzhang Shang, Leila Kokabee, Yanhui Nie, Yi Yang. Evaluating in vivo efficacy of anti-VSIG4 antibodies in humanized B-hVSIG4 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 1642.
Abstract 1644: Generation and validation of humanized GARP mice for testing novel anti-human GARP antibodies
Authors: Lyu Ruili , Li Huilin , Frame Jenna , Shen Yuelei 
Published: June 2022

Abstract
GARP is a type I transmembrane protein expressed on regulatory T cells (Tregs) and platelets. GARP expression has been shown to play an important role in the modulation of latent TGF-β1 activity by regulatory T lymphocytes (Tregs), and thus, GARP has emerged as an important regulator of peripheral immune tolerance. While GARP expression is critical for homeostasis, increased GARP expression on Tregs is associated with elevated immunosuppression in the context of tumor biology. Because of these immunomodulatory roles, targeting GARP in the context of oncologic or other diseases is of significant interest. Thus, the development of mouse models for safety and efficacy evaluation of GARP-targeted therapeutics prior to clinical trials is needed. To facilitate assessment of novel anti-human GARP antibodies, Biocytogen has generated a humanized GARP mouse model for in vitro functional validation and in vivo efficacy evaluation assays. Using CRISPR-based gene editing technology, the exons of the mouse Garp gene that encode the extracellular domain were replaced by the human GARP counterpart. mRNA Expression of the human GARP extracellular domain was confirmed in splenocytes isolated from homozygous humanized mice. Expression of human GARP was readily detected in FoxP3+ Treg cells isolated from the spleen of humanized GARP mice as assessed by flow cytometry. Importantly, the frequency of FoxP3+ Tregs and other mature leukocyte subpopulations, including T and B lymphocytes, Natural killer cells, dendritic cells, granulocytes, and monocytes/macrophages, was also similar in the blood and spleen of humanized GARP mice compared to wild-type mice. Subsequent in vivo efficacy assays using the MC38 tumor model demonstrated efficacy of the anti-human GARP antibody in slowing tumor growth in humanized GARP mice. Furthermore, combination of anti-human GARP and anti-mouse PD-1/PD-L1 blockade resulted in a significantly greater tumor inhibition than monotherapies. In summary, GARP humanized mice are a powerful platform for preclinical evaluation of anti-human GARP antibodies for cancer treatment.
Citation Format: Ruili Lyu, Huilin Li, Jenna Frame, Yuelei Shen. Generation and validation of humanized GARP mice for testing novel anti-human GARP antibodies [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 1644.
Abstract 1648: Evaluation of therapeutic drug candidates using a novel humanized B-hCSF1/hCSF1R mouse model
Authors: Lyu Ruili , Zhang Shujin , Soto Rebecca , Zhao Suman , Liu Chonghui , Yu Luke (Zhaoxue) 
Published: June 2022

Abstract
An often rate-limiting step for cancer immunotherapy is drug development. The generation of humanized mouse models can be used to assess the clinical utility of new drugs earlier in the discovery process. Tumor-associated macrophages (TAMs) represent an emerging therapeutic target, specifically high expression of the colony-stimulating factor 1 receptor (CSF1R) protein on tumors and TAMs facilitate a tumor permissive environment. Previous reports have demonstrated that blockade of the CSF1/CSF1R pathway reduced and reprogrammed intra-tumoral CSF1R+ TAMs, altogether alleviating immune suppression. To evaluate preclinical efficacy and toxicity of CSF1R targeted therapeutic candidates, Biocytogen has successfully generated a double humanized B-hCSF1/hCSF1R mouse model. The full-length encoding sequence of mouse Csf1 and the extracellular domain of Csf1r were replaced by human CSF1 and CSF1R genes, respectively. Evaluation of CSF1 and CSF1R protein expression in homozygous B-hCSF1/hCSF1R mice (H/H) was confirmed by ELISA and flow cytometry analysis. We next performed an in vivo efficacy screening study using genetically modified hCSF1 or wild-type MC38 cells inoculated into B-hCSF1/hCSF1R (H/H) mice and observed that anti-human CSF1R antibodies or combination therapy of anti-mouse PD-L1 and anti-human CSF1R antibodies were effective in controlling tumor growth. Our combined studies illustrate that Biocytogen’s novel B-hCSF1/hCSF1R mice offer a promising model for in vivo validation of CSF1 and CSF1R antibody therapeutics.
Citation Format: Ruili Lyu, Shujin Zhang, Rebecca Soto, Suman Zhao, Chonghui Liu, Luke (Zhaoxue) Yu. Evaluation of therapeutic drug candidates using a novel humanized B-hCSF1/hCSF1R mouse model [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 1648.
Abstract 165: Characterizing CLK3 in the colonic epithelium
Authors: Cabel Carly , Thorne Curtis 
Published: June 2022

Abstract
The purpose of this study is to determine the function of CLK3 in the colonic epithelium and colon cancer.
The intestinal epithelium undergoes constant renewal and is tightly regulated. Disruption of its homeostasis can result in the development of colorectal cancer (CRC). Glycogen synthase kinase 3 (GSK3) is a kinase that plays a central role in multiple signaling pathways including cellular proliferation. Our lab previously performed an siRNA screen to investigate hundreds of kinases to identify candidates which could interact with GSK3. Among the over 500 kinases that were tested, a protein kinase called CDC-like kinase 3 (CLK3) was identified as the strongest interaction with GSK3. We found that inhibition of GSK3 and CLK3 synergized to kill cancer cells. This led us to focus on CLK3 as a potential therapeutic target kinase in colon cancer.
CLK3 is part of a four-member family of CDC-like kinases involved in pre-mRNA splicing. They are dual-specificity kinases, meaning they can target both tyrosine and serine/threonine substrates. CLK3 is also a poor prognostic marker in colon cancer, with high expression of CLK3 leading to poor outcome of disease. In human colonic epithelial cells, we show that loss of CLK3 dampens Wnt response. This data, along with the strong genetic interaction with GSK3, suggests a role of CLK3 in the Wnt pathway. Because Wnt signaling is higher in the stem cell region of the crypt, and CLK3 shows a role in Wnt activity in human colonic epithelial cells, we hypothesize that CLK3 regulates mRNA splicing to promote Wnt signaling and stemness in colonic progenitor cells and carcinogenesis.
We are currently performing transcriptomics and phosphoproteomics in CLK3 CRISPR knockout cells to identify an mRNA-splicing and phosphorylation signature. We have found that the induction of Wnt-related pathway genes are lost in CLK3 knockout cells. Additionally, we are categorizing CLK3-dependent pre-mRNAs splicing events into functional groups controlling cell fate and validating in patient-derived normal and tumor organoid models. This work highlights CLK3 as a critical regulator of gut stem cells and potentially colorectal cancer stem cells.
Citation Format: Carly Cabel, Curtis Thorne. Characterizing CLK3 in the colonic epithelium [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 165.