bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3714: Role of miR-29a in pancreatic stellate cell mediated stromal remodeling in pancreatic ductal adenocarcinoma
Authors: Dey Shatovisha , Liu Sheng , Factora Tricia D. , Taleb Solaema , Riverahernandez Primavera , Udari Lata , Zhong Xiaoling , Wan Jun , Kota Janaiah 
Published: September 2020

Abstract
Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive form of malignancy with nearly equal incidence and mortality rates in patients. The disease is characterized by dense desmoplastic tumor-associated stroma, which prevents effective therapeutic delivery to the tumor core and confers resistance to chemotherapy. The central effectors of such characteristic stroma in PDAC are the pancreatic stellate cells (PSCs), which are activated by autocrine and paracrine signaling of the tumor cells and deposit excessive amounts of extracellular matrix (ECM) proteins- the major component of the stroma. This interaction of the PSCs with the cancer cells greatly promote tumor progression and metastasis. In our previous work, we demonstrated that microRNA (miRNA) 29a plays a key role in PSC-mediated stromal reaction, and that the loss of the molecule in PSCs accelerates ECM protein accumulation and cancer growth. To understand the mechanistic regulation of miR-29a in PDAC stroma, we performed RNA-seq analysis with human PSCs (hPSCs), which identified 197 genes that differentially expressed with miR-29a overexpression at a false discovery rate less than 0.05. This included 19 downregulated and one upregulated direct miR-29a targets. Of these, expression of six key targets with the highest fold changes, namely, ADAMTS2, IGF1, E2F7, LAMC1, CLDN1 and ITGA6 by miR-29a were validated at the protein level in hPSCs. In addition, canonical pathways associated with the target genes and their network interactions were assessed, which identified the association of the key targets with pathways such as collagen synthesis, insulin signaling, TGF-β and PI3K/Akt pathways. ADAMTS2, E2F7, LAMC1 and CLDN1 expressions were significantly upregulated in PDAC patient pancreas compared to that in healthy individuals. Together, the current study presents a comprehensive understanding of miR-29a mediation in stromal remodeling, and elucidates associated pathways in regulating PSCs' function in PDAC desmoplasia and pancreatic carcinogenesis.
Citation Format: Shatovisha Dey, Sheng Liu, Tricia D. Factora, Solaema Taleb, Primavera Riverahernandez, Lata Udari, Xiaoling Zhong, Jun Wan, Janaiah Kota. Role of miR-29a in pancreatic stellate cell mediated stromal remodeling in pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3714.
Abstract 3816: Elucidating the potential role of membrane complement regulatory proteins (CD46, CD55, CD59) in tumorigenesis of cervical cancer
Authors: Mo Xuetang , Leung Thomas Ho-Yin , Siu Michelle Kwan-Yee , Ngan Hextan Yuen-Sheung 
Published: September 2020

Abstract
Introduction: Cervical cancer is a common genital tract cancer. Although abundant screening and vaccination are beneficial for reducing the disease burden, the molecular mechanism underlying tumorigenicity and aggressiveness are not fully understood. Human papillomavirus (HPV) E6 protein was found to be responsible in the development of cervical cancer. Membrane complement regulatory proteins (mCRP), such as CD46, CD55, CD59, control complement activation on the surface of tumour cells and suggested to be important in contributing tumorigenesis in various types of cancers. In our recent study, we have reported that HPV E6 protein enriches the expression of CD55 in cervical cancer cells, which would further promote radio-resistance and cancer aggressiveness. In the present study, we would characterize the functional role of other two mCRPs CD46 and CD59 in contributing cervical carcinogenesis. We would further examine if triple knockout of CD46, CD55, CD59 would have synergistic effect on blockage of tumorigenesis in cervical cancer.
Material and method: Flow cytometry was performed to analyse the expression level of CD46, CD55 and CD59 in cervical cell lines C33A, C4-1, CaSki, SiHa, and a HPV18-E6-overexpressing stable clone (18E6-5, origin C33A cells). In order to investigate the effect on the blockage of those mCRPs, 18E6-5 clone with high CD46, CD55 and CD59 expression were chosen for in vitro functional study using and CRISPR/Cas9 knockout.
Results and discussion: The percentage of CD46(+), CD55(+) and CD59(+) cells were high in C4-1, CaSki, SiHa and 18E6-5 lines. The C33A cells demonstrate a low CD55(+) subpopulation while high CD46(+) and CD59(+) subpopulations. We found that single knockout of CD46 or CD59 supressed cell proliferation, sphere-forming, anchorage-independent cell growth capability, and sensitized cells to radiation treatment. In addition, triple knockout of CD46, CD55, CD59 remarkably inhibited those effects comparing with the effects of single and double knockout.
Conclusion: Silencing of CD46, CD55, CD59 expression abolished the tumorigenic, radio-resistant and aggressive effects in cervical cancer cells, with triple knockout displayed the highest blockage effects. Those mCRPs would be responsible for tumorigenicity, radio-resistance and cancer aggressiveness in cervical cancer.
Citation Format: Xuetang Mo, Thomas Ho-Yin Leung, Michelle Kwan-Yee Siu, Hextan Yuen-Sheung Ngan. Elucidating the potential role of membrane complement regulatory proteins (CD46, CD55, CD59) in tumorigenesis of cervical cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3816.
Abstract 3822: Genome-wide CRISPR library screen to identify synthetic lethal targets of a WEE1 degrader
Authors: Yue Yangbo , Su Hexiu 
Published: September 2020

Abstract
Background: WEE1 kinase is a G2/M cell cycle checkpoint protein, activated to allow DNA repair prior to cell entry to mitosis. WEE1 kinase inhibitors have witnessed encouraging early clinical results in combination with chemotherapy and radiotherapy. We previously identified a small molecule WEE1 degrader and showed selective growth inhibition in pancreatic cancer cell lines. To further study the functional mechanism of this compound and to investigate the combinational inhibition of WEE1 degradation, we performed genome-wide CRISPR library screening in pancreatic cancer cell lines harboring either wildtype K-RAS or mutant K-RAS. The identified synthetic lethal targets may lead to tractable therapeutic opportunities in combination with this compound.
Method: We conducted CRISPR screening in MIA-PaCa2 and PANC1 cell lines, harboring mutant and wild type K-RAS, respectively, with genome-wide Brunello library (2 vector system) in the presence or absence of the WEE1 degrader for 2 weeks. After harvesting the cells for deep sequencing, we analyzed the top hits whose inhibition showed synergetic cytotoxic effect with the compound.
Results: According to the sequencing result, the deletion of several candidate genes increased the sensitivity to the compound, indicating combinational potential with this WEE1 degrader. Further validation studies are in progress. We will discuss possible mechanisms underlying the synthetic lethality. The pipeline we have established, from experimental design to bioinformatics analysis, can be applied to investigate functional mechanism and combination strategy of any target therapeutic agents.
Citation Format: Yangbo Yue, Hexiu Su. Genome-wide CRISPR library screen to identify synthetic lethal targets of a WEE1 degrader [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3822.
Abstract 387: Analysis of FFPE human tumor tissues using CODEX with signal amplification
Authors: Mistry Sejal , Dakshinamoorthy Gajalakshmi , Yuan Jessica , Noordam Pieter , Kim Joseph , Lee Won-Mean , Kennedy-Darling Julia 
Published: September 2020

Abstract
Analysis of FFPE human tumor tissues using CODEX with signal amplification
Characterizing the complexities of the tumor microenvironment is fundamental to understanding disease mechanisms. The spatial relationships between infiltrating immune cells and the remodeling of the cellular matrix is widely recognized as a key component to defining tumor heterogeneity. Current methodologies for studying cells within the context of tissue architecture, like traditional immunofluorescence (IF) and immunohistochemistry (IHC), are limiting–allowing the assessment of only a few parameters at a time. The CODEX technology has overcome this limitation through a DNA-based labeling strategy, involving adding and removing dye-labeled oligonucleotides (reporters) across multiple cycles to oligonucleotide-labeled antibodies. In this manner, tens of markers can be analyzed on the same tissue. Additionally, CODEX interfaces with existing inverted microscopes and provides a cost-effective, fully automated platform for highly multiplexed imaging. Recently, we expanded our workflow to amplify the fluorescent signal intensities of low expressing biomarkers by combining Tyramide Signal Amplification (TSA) with CODEX. This approach is associated with key markers used to analyze the tumor microenvironment, including PD-L1 and PD-1. We demonstrated signal amplification of more than 50X compared with non-amplified signal. Here, we present the analysis of CODEX data for a panel of over 20 biomarkers on human FFPE tissue including signal amplification of PD-L1, PD-1 and foxp3. Through amplification of key markers as the final cycle of a CODEX experiment, low expression can be measured and used to identify key cell types, including Treg cells and PD-L1 positive tumor and stromal regions.
Citation Format: Sejal Mistry, Gajalakshmi Dakshinamoorthy, Jessica Yuan, Pieter Noordam, Joseph Kim, Won-Mean Lee, Julia Kennedy-Darling. Analysis of FFPE human tumor tissues using CODEX with signal amplification [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 387.
Abstract 3871: Infiltration of CD56dim NK cells regulated by MLH1 into tumor tissue of FAP patients predict better prognosis
Authors: Yang Jun , Zhang Hushan , Liu Wei Qing , Li Wenliang , Ma Shumin , Liang Lei , Kou Zhiyong , Huang Depei , Shen Xudong , Dong Jian 
Published: September 2020

Abstract
Backgroud
Familial adenomatous polyposis (FAP) is an autosomal dominant-inherited colorectal cancer (CRC) with poor prognosis, and is also considered as model to study the change of CRC from pre-tumor to tumor progression. To better predict and guide clinical applications of immune check point inhibitors(ICI), researchers proposed the classification system of human cancer, termed tumor immunity in the microenvironment (TIME), that four different TIME groups have been identified according to the presence of TILs and B7-H1(PD-L1) expression in tumor biopsies. Therefore, we followed and performed this theory into our research about TIME of FAP, to explore the potential of immunotherapy for FAP, and explore the cancerous mechanism from immune microenvironment.
Methods
We collected tissue samples of 40 FAP patients from May 2009 to December 2018, followed by detection of PD-L1 by using Dako PD-L1 IHC 22C3 pharmDx, MLH1, PMS2, MSH2, MSH6 were detected by IHC, and CD8+ T cells, tumor associated macrophages (TAM) and NK cells infiltration in the tumor biopsies by multiple fluorescence immunohistochemistry (mICH). Follow-up data of these 40 patients were analyzed.
Results
CD8+T cells, M1 and M2 TAM, CD56bright NK and CD56dim NK cells were found both in intratumoral region and tumor rim of FAP patients(n=40), no differences have been found between M1and M2 TAM, however, CD56dim NK increased more than CD56bright NK both in intratumoral region and tumor rim(p&lt;0.01). Surprisingly, no positive expression of PD-L1 was detected in the tumor tissue of these FAP patients (Tumor Proportion Score, TPS&lt;1%). Further results shown CD56dim NK mainly increased in the tissue of FAP patients(n=7) with loss of MLH1, and consistent with this, these patients shown better prognosis than patients with wild type MLH1, that is, have longer progression free survival (p&lt;0.005).
Conclusions
For the TIME characteristics of FAP patient, Immunotherapy targeted PD-1/PD-L1 may not applicable for FAP, however, MLH1, the member of mismatch repair protein, was associated with infiltration of NK cells, especially CD56dim NK cells in the tumor, and also associated with prognosis of FAP patients. Therefore, it can be exploited as potential target of immunotherapy for FAP or even CRC.
Citation Format: Jun Yang, Hushan Zhang, Wei Qing Liu, Wenliang Li, Shumin Ma, Lei Liang, Zhiyong Kou, Depei Huang, Xudong Shen, Jian Dong. Infiltration of CD56dim NK cells regulated by MLH1 into tumor tissue of FAP patients predict better prognosis [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3871.
Abstract 3911: High-throughput screening of patient-derived HCC organoids for drug discovery
Authors: Wong Alissa Michelle Go , Cheung Winnie , Wong Maggie , Wong Aikha , Lam Helena , Zhang Zhe , Man Matthew , Chen Hui , Close David , Johnston Paul , Chan Anthony , Lai Paul , Kwong Joseph , Wong Nathalie 
Published: September 2020

Abstract
Current in vitro models of primary liver cancer insufficiently recapitulates the tumor heterogeneity and pathophysiology of the original patient's tumor. This significantly hinders translational studies and often leads to the high rate of clinical trial attrition and poor patient response. Recent technological advances in organoid culture methods have been reported to overcome these limitations. Our group has successfully established 3 patient-derived HCC tumor organoids that closely recapitulate the patient's pathophysiology. Conventional short-term monolayer culture of the same 3 tumors as well as 3 non-tumoral adjacent liver and 3 normal liver organoids were established and compared with 5 HCC cell lines from ATCC for their drug response profiles. Subsequent drug screening was performed using a panel of 100 drugs, of which 70 are FDA-approved for either HCC or other cancers, as well as 30 pipeline compounds targeting frequently mutated pathways in HCC. We found that our screening pipeline could achieve robust z' scores and reproducible data points with technical and biological replicates. Principal component analysis of Total Area under curve (AUC) values show distinct and progressive changes between Normal Liver, Non-tumoral adjacent liver, and Tumor organoids. Upon comparison with HCC cell lines Hep3B, HepG2, Huh7, SNU398 and PLC5, we found HCC patient-derived short term cultures, particularly organoids have more diverse drug sensitivity profiles and may be a better indicator of patient heterogeneity. Interestingly, FDA-approved drugs for HCC such as Regorafenib show high sensitivity and tumor specificity to ATCC cell lines (ATCC vs Normal liver P=0.00017***), but this specificity was reduced and lost in patient-derived 2D cultures (P=0.045*) and organoids (P=0.895), respectively. This may explain the observed hepatotoxicity observed in HCC patients and warrant further investigation of more targeted ways of treating HCC.
Citation Format: Alissa Michelle Go Wong, Winnie Cheung, Maggie Wong, Aikha Wong, Helena Lam, Zhe Zhang, Matthew Man, Hui Chen, David Close, Paul Johnston, Anthony Chan, Paul Lai, Joseph Kwong, Nathalie Wong. High-throughput screening of patient-derived HCC organoids for drug discovery [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3911.
Abstract 394: LXR agonist N,N-dimethyl-3-β-hydroxy-cholenamide, DMHCA, reduces ErbB2-dependent tumorigenesis and immune tolerance in a conditional model of mammary fibrosis
Authors: Glazer Robert I. , Sheng Gao , Yuan Hongyan , Ranjit Suman , Lu Jin , Lu Xun , Levi Moshe 
Published: September 2020

Abstract
Background: One of the central challenges for cancer therapy is the identification of factors in the tumor microenvironment that increase tumor progression and prevent the immune system from eradicating the tumor. One such factor associated with breast cancer is stromal fibrosis, a histopathologic criterion for invasive cancer and poor survival. Fibrosis is caused by inflammatory factors and remodeling of the extracellular matrix, which results in excessive collagen deposition and an immune tolerant environment. Experimental Procedures: We developed a novel transgenic model representative of HER2/ErbB2+ breast cancer, termed NeuT/ATTAC, in which partial ablation of mammary fat resulted in its replacement with fibrotic tissue. NeuT/ATTAC mice were administered a diet supplemented with 0.04% DMHCA, a desmosterol-like pan agonist of the liver X receptor (LXR). Collagen deposition resulting from fibrosis was assessed by second harmonic generation (SHG) fluorescence microscopy and histological staining, immune cell subsets were analyzed by fluorescence-activated cell sorting and biomarkers of fibrosis, proliferation and immune tolerance were determined by IHC. Findings: Administration of DMHCA to NeuT/ATTAC mice beginning at two months of age reduced tumorigenesis, tumor multiplicity and suppressed fibrosis as shown by loss of collagen, fibroblast activation protein (FAP) and α-smooth muscle actin (SMA). These changes were accompanied by a marked reduction in the percentage of MDSC, and an increase in CD8+ effector T cells and PD1+/Foxp3+ CD4 T cells.
Conclusion: Reduction of fibrosis, tumorigenesis and immune tolerance by DMHCA suggests that this LXR agonist may be useful for reducing tumor-associated fibrosis and in combination with immune checkpoint inhibitors that target PD1+ Treg cells for moderating immune tolerance. Unlike non-steroidal LXR agonists, DMHCA does not increase serum triglycerides or produce hepatic steatosis, and may therefore be a good candidate for clinical studies. Supported by a Sher Award, 1P30CA051008 from the NCI, NIH, to the Lombardi Comprehensive Cancer Center, and R01 HL133545 and R01 DK116567 from the NIH to M.L.
Citation Format: Robert I. Glazer, Gao Sheng, Hongyan Yuan, Suman Ranjit, Jin Lu, Xun Lu, Moshe Levi. LXR agonist N,N-dimethyl-3-β-hydroxy-cholenamide, DMHCA, reduces ErbB2-dependent tumorigenesis and immune tolerance in a conditional model of mammary fibrosis [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 394.
Abstract 3965: Novel biomimetic, interfacing 3D hydrogels to investigate roles of glioblastoma (GBM) niche on GBM progression
Authors: Sohrabi Alireza , Liang Jesse , Safarians Gevick , Guivatchian Elnaz , Solomon Itay , Seidlits Stephanie 
Published: September 2020

Abstract
Glioblastoma (GBM) is the most common and lethal form of brain tumors with survival rate of 5% after 5 years. In the tumor microenvironment, invading GBM cells experience various ECM transitions at the tumor margin, such as alterations in matrix stiffness. Although conventional three-dimensional (3D) hydrogels have improved upon existing in vitro tumor models, these biomaterial platforms are not capable of separating the migratory GBM population from the bulk of the tumor. In addition, conventional hydrogels can only mimic a single microenvironment property (i.e. stiffness and protein presentation). In this study, we fabricated interfacing hyaluronic acid (HA)-based hydrogels using thiol-ene photochemistry to mimic the distinct stiffness conditions of tumor and surrounding tissue as well as the stiffness transition. The novel interfacing model enabled us to isolate invading GBM cells for further in vitro analysis. Atomic Force Microscopy (AFM) analysis of our xenograft GBM tumors in mice brain demonstrated that GBM tumor is stiffer than surrounding brain tissue. Therefore, mechanical properties of each hydrogel condition were characterized via AFM to confirm similarity of hydrogel stiffness to GBM xenografts and patient-derived, primary GBM cell migration in the HA-hydrogels was investigated. Immunohistochemistry staining showed that invading GBM cells, passing the interface have higher expression of RHAMM and CD44. Our findings suggest that invading GBM cells might have different expression profile and therefore require more thorough analyses using single cell RNA-seq.
Citation Format: Alireza Sohrabi, Jesse Liang, Gevick Safarians, Elnaz Guivatchian, Itay Solomon, Stephanie Seidlits. Novel biomimetic, interfacing 3D hydrogels to investigate roles of glioblastoma (GBM) niche on GBM progression [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3965.