bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 600: Discovery of a single-domain antibody fragment targeting PD-L1 for cancer immunotherapy
Authors: Liu Yanli , Liu Hao , Cheng Kun 
Published: June 2022

Abstract
Immunotherapy has shown enormous promise for cancer treatment. The use of Antibodies to target immune checkpoints, particularly PD-1/PD-L1, has made a profound impact in cancer immunotherapy. Single-domain antibody fragment (sdAb) is an antibody fragment composed of a single variable domain of the heavy chain. sdAb is the smallest antibody fragment (15 kDa) that maintains the similar antigen-binding affinity. sdAbs show many advantages over monoclonal antibodies such as small size, low immunogenicity, ease of production, and ease of modification, making sdAbs promising candidates for cancer immunotherapy. Here, we use a phage display library to identify an anti-PD-L1 sdAb (CLV3) which effectively blocks the PD-1/PD-L1 interaction. The CLV3 sdAb demonstrates high binding affinity and specificity to PD-L1 protein and PD-L1 overexpression DU145 tumor cells. Moreover, the CLV3 sdAb exhibits efficient tumor penetration in a 3D tumor spheroid model, suggesting its good penetration capability in the tumor microenvironment in vivo. CLV3 restores T cell proliferation and reduces T cell apoptosis in the co-culture experiment of peripheral blood mononuclear cells (PBMC) with tumor cells. These data suggest that the CLV3 sdAb can be potentially used as an anti-PD-L1 inhibitor for cancer immunotherapy.
Citation Format: Yanli Liu, Hao Liu, Kun Cheng. Discovery of a single-domain antibody fragment targeting PD-L1 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 600.
Abstract 6007: Identification of drivers of soft tissue sarcoma metastasis
Authors: Carr-Ascher Janai R. , Munoz Maria 
Published: June 2022

Abstract
Soft tissue sarcomas are a rare and heterogeneous group of tumors with a 65% five-year survival rate despite aggressive treatment with surgery, radiation, and chemotherapy. Even with intensive multimodality treatment, many patients will develop incurable metastatic disease, providing evidence that our current therapies are ineffective and new treatments are desperately needed. Pre-clinical models to study metastasis of human soft tissue sarcomas are limited. We have developed a system that allows for the formation of distinct soft tissue sarcoma subtypes from mesenchymal stem cells that have been depleted of p53 and Rb1 using CRISPR. Injection of these cells intramuscularly results in metastasis to the lung. Using this novel model system, we have performed a genetic screen to identify pathways and specific drivers of sarcoma metastasis. This pre-clinical in vivo model system will allow for the identification of putative therapeutic targets to prevent the development of lethal metastatic growth.
Citation Format: Janai R. Carr-Ascher, Maria Munoz. Identification of drivers of soft tissue sarcoma metastasis [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 6007.
Abstract 6011: RNA-seq analysis of liver from NASH derived HCC model mouse treated with streptozotocin-high fat diet
Authors: Hashiguchi Taishi , Linh Bui Phuong , Sakakibara Yuki , Tanaka Ryuto , Pigney Elizabeth H 
Published: June 2022

Abstract
Introduction; Non-alcoholic steatohepatitis (NASH) is a chronic liver disease often associated with type 2 diabetes. It sometimes progresses to more serious conditions such as liver fibrosis and hepatocellular carcinoma (HCC). Therapeutic agents for NASH have not yet been developed, and an animal model with high clinical correlation is required. The STAM mouse is a NASH model mouse with a background of type 2 diabetes. This mouse model shows the same pathological progression as human NASH patients, and has been used for the evaluation of many drug candidates, as well as basic research. However the gene profile, including mutations, has not been elucidated in this model. In this study, we analyzed the RNA-seq data of STAM mice at each pathological stage and examined the clinical correlation at the gene level.
Methods;NASH was induced in male mice by a single subcutaneous injection of 200 ug streptozotocin solution 2 days after birth and feeding with high fat diet after 4 weeks of age. The mice were sacrificed and livers collected at 6, 8, 10, 12, 16 and 20 weeks of age. For liver samples, the left lateral lobe was snap frozen in liquid nitrogen and stored at -80°C for RNA-seq analysis. Total RNA of the cells was isolated using RNeasy mini kit.
Results: The gene expression of the canonical pathway in NASH progression from steatosis to hepatocellular carcinoma was analyzed. Increased expression was observed in CCL2, TLR4, HSP47, alpha-SMA, c-Myc, and cyclin D1. Since it has been reported that genetic traits are involved in the development of NASH-HCC, we next analyzed the genetic mutations in the STAM mice. The number of individuals showing mutations in Mtorinvolved in Insulin signaling increases as the disease progresses, especially in the liver cancer phase.
Citation Format: Taishi Hashiguchi, Bui Phuong Linh, Yuki Sakakibara, Ryuto Tanaka, Elizabeth H Pigney. RNA-seq analysis of liver from NASH derived HCC model mouse treated with streptozotocin-high fat diet [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 6011.
Abstract 6015: Comprehensive evaluation of three-dimensional (3D) culture systems as compound screening platforms with primary cancer cell lines
Authors: Zhang Chao , Wu Qinghua , Jin Lei , Chen Deyin , Tang Xuzhen , Qi Hui , Gao Wenjie , Ji Qunsheng 
Published: June 2022

Abstract
There are a number of challenges associated with personalized medicines. For example, many models revolve around the question of whether or not the in vitro models capture the relevant in vivo pathobiology and drug response information that may impact a patient's response to a therapy. Most in vitro oncology drug screening techniques are based on two-dimensional (2D) cell culture, which, to date, has failed to accurately imitate the physiological microenvironments due to the inherent limitations, such as the lack of interactions between intracellular and extracellular environments, or modifications in three-dimensional cell morphology, polarity, and division. These disadvantages prompted the development of models which would better simulate the microenvironment of tumor cells. Three-dimensional (3D) culture systems are becoming increasingly popular due to their ability to mimic natural cell characteristics and tissue-like structures compared with traditional 2D monolayer cell culture. Currently, multicellular spheroid and organoid cell culture are two popular 3D culture methods to evaluate drug activity. Spheroid cell culture utilizes ultra-low attachment plate to allow cells grow in spheroid format. Organoid cell culture is accomplished by growing cells in extracellular matrix scaffolds (such as Matrigel). In this study, we attempted to compare different drug screening platforms, including patient-derived xenograft (PDX) and two 3D cell cultures (spheroid and organoid). Compound sensitivity, genomics, pathology and expression of clinical marker were profiled. The purpose of this study is to identify an optimal model as a drug testing platform and a bridge between in vitro studies and animal experiments for identifying novel therapeutic targets.
Citation Format: Chao Zhang, Qinghua Wu, Lei Jin, Deyin Chen, Xuzhen Tang, Hui Qi, Wenjie Gao, Qunsheng Ji. Comprehensive evaluation of three-dimensional (3D) culture systems as compound screening platforms with primary cancer cell lines [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 6015.
Abstract 6028: Patient derived organoids demonstrate hallmark characteristics of small cell lung cancer
Authors: D'Rozario Joshua , George Julie , Thomas Roman 
Published: June 2022

Abstract
Small cell lung cancer (SCLC) accounts for approximately 15% of all lung cancers with an overall 5-year survival rate of 5-10%. Although transgenic mouse models and patient derived xenograft models have been shown to capture the heterogeneity and complexity of SCLC, the costs involved and timeframe of generating these models can be extensive. Furthermore, these models lack the ability to study the patients’ immune components and tumor microenvironment. Alternatively, in other cancer types in vitro methods to generate organoids have been successful in the hope to develop personalized patient derived organoids for screening and further downstream analyses. Here, we have generated and characterized small cell lung cancer patient-derived tumor organoids and confirmed that the organoids possess similar molecular characteristics, genetic profiles and morphological architectures to the corresponding patient tumor tissue. Organoids were shown to express hallmark features for SCLC, which included expression of neuroendocrine markers CD56, CHGA, INSM1 and SYP shown by immunohistochemistry. We suggest utilizing these organoids as SCLC patient models would be ideal for investigating the tumor microenvironment in SCLC, mechanisms between tumor immune cell interactions as well as assessing patients’ responses to therapy.
Citation Format: Joshua D'Rozario, Julie George, Roman Thomas. Patient derived organoids demonstrate hallmark characteristics of small cell lung 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 6028.
Abstract 6088: Reconstructing mutational lineages in breast cancer by multi-patient-targeted single cell DNA sequencing
Authors: Leighton Jake , Hu Min , Sei Emi , Meric-Bernstam Funda , Navin Nicholas 
Published: June 2022

Abstract
Single cell DNA sequencing (scDNA-seq) methods are powerful tools for profiling mutations in cancer cells, however most genomic regions characterized in single cells are non-informative. To overcome this issue, we developed a Multi-Patient-Targeted (MPT) scDNA-seq sequencing method. MPT involves first performing bulk exome sequencing across a cohort of cancer patients to identify somatic mutations, which are then pooled together to develop a single custom targeted panel for high-throughput scDNA-seq using a microfluidics platform. We applied MPT to profile 330 mutations across 23,500 cells from 5 TNBC patients, which showed that 3 tumors were monoclonal and 2 tumors were polyclonal. From this data, we reconstructed mutational lineages and identified early mutational and copy number events, including early TP53 mutations that occurred in all five patients. Collectively, our data suggests that MPT can overcome technical obstacles for studying tumor evolution using scDNA-seq by profiling information-rich mutation sites.
Citation Format: Jake Leighton, Min Hu, Emi Sei, Funda Meric-Bernstam, Nicholas Navin. Reconstructing mutational lineages in breast cancer by multi-patient-targeted single cell DNA sequencing [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 6088.
Abstract 6100: Chemotherapy reduces TGF-beta signaling triggered by blockade of the PD-1/PD-L1 axis
Authors: Iida Masafumi , Horn Lucas A , Qin Haiyan , Fousek Kristen , Schlom Jeffrey , Palena Claudia 
Published: June 2022

Abstract
Combination of immune checkpoint blockade (ICB) with chemotherapy is becoming more commonly used for the treatment of various cancer types, including advanced non-small cell lung cancer, and triple negative breast cancer (TNBC) in patients who have PD-L1 positive tumors, among others. Although chemotherapy-ICB modalities have been shown to improve clinical responses over those observed with ICB alone, long-term disease control is achieved only in a small subset of patients. To provide a rational basis for combination therapy, and to potentially develop more effective approaches, in this study we investigated the impact of chemotherapy on the tumor microenvironment (TME) in the context of PD-1/PD-L1 axis blockade. ICB refractory murine models were utilized, including the TNBC 4T1 model and the Lewis Lung Carcinoma (LLC) model. Following treatments with anti-PD-L1, docetaxel, or the combination anti-PD-L1 plus docetaxel, tumors were collected 4 days after the last treatment to evaluate early changes in the TME. Analyses included the characterization of tumor immune infiltrates via flow cytometry, RNA expression profiling via RT-PCR, RNA in situ hybridization, and immunohistochemistry for detection of immune infiltrates, cancer-associated fibroblasts, and phospho-Smad2 signaling. RNA analysis of 4T1 and LLC tumors revealed a significant upregulation of several transforming growth factor-beta (TGF-beta) downstream target genes in the anti-PD-L1 monotherapy group. This increase was associated with enhanced expression of TGF-beta in the TME, as determined by RNA in situ hybridization, together with enhanced TGF-beta signaling denoted by increased expression of phospho-Smad2. Tumors treated with anti-PD-L1 monotherapy also exhibited increased numbers of alpha-smooth muscle actin positive fibroblasts in the TME. The effects observed with anti-PD-L1 as a monotherapy were not observed in tumors treated with docetaxel monotherapy or in those treated with the combination docetaxel plus anti-PD-L1 therapy, suggesting that upregulation of immunosuppressive TGF-beta signaling in the TME could be prevented by the addition of chemotherapy. Ongoing and future studies are aimed at understanding the mechanisms involved in the upregulation of TGF-beta signaling in the TME in response to anti-PD-L1 therapy in non-responsive tumors, the effect of various chemotherapies in this phenomenon, and the study of additional tumor models for optimization of chemotherapy-ICB modalities.
Citation Format: Masafumi Iida, Lucas A Horn, Haiyan Qin, Kristen Fousek, Jeffrey Schlom, Claudia Palena. Chemotherapy reduces TGF-beta signaling triggered by blockade of the PD-1/PD-L1 axis [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 6100.
Abstract 6102: Single-cell RNA sequencing of lung adenocarcinoma patients reveals age-dependent tumour microenvironment alterations that facilitate response to immunotherapy
Authors: Zhu Xiaoqiang , Wang Hai , Pan Xuefeng , Hong Jie , Wong Jason Wing Hon , Chen Haoyan , Zhang Youwei 
Published: June 2022

Abstract
Background: Immunotherapy has revolutionized the treatment of lung adenocarcinoma (LUAD). While the increase in cancer incidence with age and age-associated changes in immune function have been well characterized, age-related differences in the intratumoral immune populations and how this affects immunotherapy response remain unknown.
Methods: We collected clinical data from 3,544 samples treated with immune checkpoint blockade (ICB) at the pan-cancer level. Single-cell RNA sequencing data from 19 LUAD samples were integrated and analyzed. Cell types were identified based on canonical marker gene expression.
Results: Aged individuals responded more efficiently to ICB treatment at the pan-cancer level. By focusing on LUAD, we showed that patients aged above 46 responded better and had longer progression-free survival than younger counterparts. Further, we presented a 58,031-cell catalog of age-associated single-cell transcriptomes from LUAD patients. Our analysis revealed the accumulation of immune response and chronic inflammation with increasing age. Aged tissues (&gt; 60 years) displayed dominant exhausted natural killer (NK)/T cells and T regulatory cells and were enriched with two subtypes of tumour-associated macrophages marked by SPP1 and CX3CR1 expression, respectively. Furthermore, the ageing dynamics of stroma cells created a pro-tumor microenvironment reflected by increased angiogenesis and collagen formation.
Conclusions: These results revealed an increasing immune-suppressive tumour microenvironment with age, supporting an observation of improved ICB response in older LUAD patients. Our extensive single-cell analysis enhances the understanding of age-dependent molecular and cellular dynamics in LUAD. Our results highlight the importance of considering age as a factor for ICB response and provide a rich resource for developing therapeutic targets in LUAD-microenvironment interactions.
Citation Format: Xiaoqiang Zhu, Hai Wang, Xuefeng Pan, Jie Hong, Jason Wing Hon Wong, Haoyan Chen, Youwei Zhang. Single-cell RNA sequencing of lung adenocarcinoma patients reveals age-dependent tumour microenvironment alterations that facilitate response to 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 6102.
Abstract 6191: A novel anti-IL33 antibody that neutralized Th1-type and Th2-type pro-inflammation responses induced by IL-33/ST2 axis pathway
Authors: Zhang Xuesai , Li Qingrou , Liu Cui , Zhao Le , Wang Lujie , Deng Lan , Xu Wei , song Hongli , Zhu Zhenping , Huang Haomin 
Published: June 2022

Abstract
IL-33, the ligand for ST2, is a member of IL-1 family. IL-33 was described to activate many immune cell types involved in type-2 immunity and allergic inflammation. Here we describe a novel anti-IL33 humanized monoclonal antibody, SSGJ-621, which bound IL-33 with high affinity and blocked the interaction between IL-33 and ST2. Our study has shown that SSGJ-621 was capable of effectively neutralizing IL-33-induced IL-6 secretion from HUVECs and IL-33-dependent IFNγ production in PBMCs and NK cells, suggesting a role that IL-33 might be involved in Th1-type of immunity. In addition, SSGJ-621 also exhibited potent inhibitory effects on IL-33 induced IL-5 and IL-13 production in KU812 cells. In vivo study demonstrated that SSGJ-621 can inhibit the activation of splenocytes in spleen and the production of eosinophils and IL-5 in peripheral blood. Thereby, our results highlight the dual functions of IL-33 in promoting Th1 and Th2 immune responses and SSGJ-621 has potentials to treat not only chronic inflammatory diseases induced by IL-33, but also cancers by favoring Th1 antitumor immunity in the tumor microenvironment.
Citation Format: Xuesai Zhang, Qingrou Li, Cui Liu, Le Zhao, Lujie Wang, Lan Deng, Wei Xu, Hongli song, Zhenping Zhu, Haomin Huang. A novel anti-IL33 antibody that neutralized Th1-type and Th2-type pro-inflammation responses induced by IL-33/ST2 axis pathway [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 6191.
Abstract 6197: The role of the integrated stress response in pancreatic cancer adaptation to tumor microenvironmental nutrient stress
Authors: Cai Chufan 
Published: June 2022

Abstract
Pancreatic ductal adenocarcinoma (PDAC) is one of the deadliest cancers, with a five-year survival rate ~10%1. PDAC is characterized by an altered/abnormal nutrient tumormicroenvironment (TME)2, and cancer cells must adapt metabolically, which exposes potentialtherapeutic targetable vulnerabilities. Therefore, we seek to identify metabolic pathways thatallow PDAC adaptation to such abnormal TME. The integrated stress response (ISR) is anintracellular stress response pathway that senses diverse stressors, and subsequently initiatesdownstream stress adaptations3. We find both murine and human PDAC have elevatedexpression of ATF4, the most well-characterized ISR effector, compared to paired healthypancreas by immunohistochemistry staining. Further, I find the ISR is activated and critical forPDAC survival/proliferation upon physiological PDAC TME nutrient stress. What is triggering theISR activation in PDAC? Through a variety of studies, I determined that it was the amino acidstress, specifically the depletion of arginine and asparagine, that activated the ISR upstreamkinase GCN2, which is responsible for ATF4 upregulation upon PDAC TME nutritional stress.Through CRISPR-based competition assays, I determined that ATF4 activity was critical forPDAC cells to cope with the depletion of arginine and asparagine, and ATF4 activity wasdispensable when these two amino acids were supplied even though cultured in PDACphysiological TME conditions. Currently, I am seeking to validate these findings in murine PDACmodels, and to determine if ATF4 or GCN2 could be relevant therapeutic targets, using an invivo tumor competition assay that I am developing. Altogether, I find the ISR is active in PDACboth ex vivo and in vivo, and suggest that the ISR is critical for PDAC survival/proliferation byallowing adaptation to abnormal PDAC amino acid TME. This work suggests that the ISR is anovel therapeutic target for PDAC treatment, which ultimately can benefit PDAC patients.
References: 1. Cancer Facts & Figures 2021. https://www.cancer.org/research/cancer-facts-statistics/all-cancer-facts-figures/cancer-facts-figures-2021.html. 2. Sullivan, M. R. et al. Quantification of microenvironmental metabolites in murine cancersreveals determinants of tumor nutrient availability. Elife 8, (2019).3. Pakos-Zebrucka, K. et al. The integrated stress response. EMBO Rep. 17, 1374-1395(2016).
Citation Format: Chufan Cai. The role of the integrated stress response in pancreatic cancer adaptation to tumor microenvironmental nutrient stress [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 6197.