bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 5548: Indicators of immune activation upon p14ARF and interferon-beta gene transfer in a human melanoma cell line
Authors: Cerqueira Otto Luiz , Salomon Maria Alejandra , Cardoso Elaine Cristina , Tortelli Tharcisio Citrangulo , Strauss Bryan Eric 
Published: September 2020

Abstract
Immunoevasion adopted by tumor cells is an important cancer hallmark and understanding of its mechanisms has generated promising therapeutic approaches. In this context, triggering immunogenic cell death (ICD) is a promising idea since it alarms the immune system by attracting a myriad of cell populations that promote innate and adaptive immune responses. Since melanomas preserve wild-type p53 in over 80% of cases, our group developed a gene therapy strategy that utilizes an adenoviral vector with a p53-responsive promoter (PG). Here we present a critical advance for our approach, examining the immune activating potential of combined p14ARF and IFN-beta gene transfer in human melanoma cells. Our results indicate that IFN-beta alone or in combination with p14ARF was able to induce massive cell death accompanied by caspase 3/7 activation in the human melanoma cell line SKMEL147. In our animal model of in situ gene therapy, s.c. SKMEL147 tumors were established in Nod-scid mice before intratumoral virus injection, revealing tumor inhibition and improved survival in response to IFN-beta alone or in combination with p14ARF. However, this model cannot reveal immune activation and so far the benefit of p14ARF has not been revealed. Strikingly, three markers of ICD (exposition of calrecticulin, secretion of ATP and IFN-beta) were induced only when cells were treated with combined p14ARF and IFN-beta gene transfer. We then used PBMCs derived from normal donors as a source of DCs and lymphocytes. Coculture of DCs with previously-transduced SKMEL147 exhibit a typical phenotype of activation. Finally, coculture of these exposed DCs with lymphocytes results in high-level secretion of proinflammatory cytokines, followed by the increase of IL-10 in the samples treated with combination of p14ARF and IFN-beta. Collectively, our results indicate that the advantage of p14ARF and INF-beta delivered by our adenoviral system is the induction of cell death in human melanoma cells by an immune-activating mechanism. Future studies will include examination of this phenomenon in tumors from melanoma patients and using their own PBMCS. In parallel, the use of PDX model for in situ treatment will also be explored.
Citation Format: Otto Luiz Cerqueira, Maria Alejandra Salomon, Elaine Cristina Cardoso, Tharcisio Citrangulo Tortelli, Bryan Eric Strauss. Indicators of immune activation upon p14ARF and interferon-beta gene transfer in a human melanoma cell line [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 5548.
Abstract 5607: Broad acting glutamine antagonism remodels the tumor microenvironment, induces distinctive immune modulation, and synergizes with immune checkpoint blockade
Authors: Yokoyama Yumi , Wild Robert 
Published: September 2020

Abstract
We previously reported that DRP-104, a novel broad acting glutamine antagonist, has significant therapeutic potential in cancer via directly targeting tumor metabolism and inducing a potent antitumor immune response. Here we sought to further elucidate the immunomodulatory effect of DRP-104 on tumor growth inhibition as a single agent and in combination with PD-1/PD-L1 checkpoint inhibitors in the MC38 and CT26 colon cancer syngeneic models. We utilized multiple technologies to assess mechanistic effects on immune cells in vivoincluding flowcytometry, multiplex immunoassay, gene expression profiling and GeoMx™ digital spatial profiling. DRP-104 mediated tumor growth inhibition was associated with increased tumor-infiltrating leukocytes (TIL) including T, NKT, and NK cells, M1-polarized tumor associated macrophages, and decreased immune-suppressive cells such as MDSCs. Nanostring® IO360 analysis revealed broad immunological modulation such as increase in cytotoxicity/antigen presentation score, increase in tumor metabolic stress/apoptotic score and decrease in cell proliferation score. GeoMx™ profiling also showed increased tumoral T cells and granzyme B expression. Luminex analysis of tumor lysates revealed DRP-104 treatment decreased pro-tumorigenic proteins, such as VEGF and IL-8. DRP-104 showed significant dose dependent tumor growth inhibition and regressions as a single agent in mouse syngeneic tumor models, including those resistant to immune checkpoint inhibitors. Lastly, combination of DRP-104 with anti-PD-1 Ab or anti-PD-L1 Ab further reduced tumor growth resulting in significant improvement in survival with long-term durable cures. In summary, DRP-104 treatment resulted in broad remodeling of the tumor microenvironment including enhanced infiltration and function of multiple immune cells distinct from activities obtained by a checkpoint inhibitor treatment. Combination therapy of DRP-104 with anti-PD-1/PD-L1 achieved significantly enhanced anti-tumor efficacy including long-term durable cures even in checkpoint inhibitor resistant models. This unique and non-overlapping mechanism of action supports clinical development of DRP-104 alone and in combination with PD-1/PD-L-1 checkpoint inhibitors.
Citation Format: Yumi Yokoyama, Robert Wild. Broad acting glutamine antagonism remodels the tumor microenvironment, induces distinctive immune modulation, and synergizes with immune checkpoint blockade [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 5607.
Abstract 5613: Human interleukin 15 (IL15) humanized NCG mice support the human natural killer cells reconstitution and development
Authors: Ju Cunxiang , Zhang Mingkun , Wu Dan , Wang Dingyu , Sun Hongyan , Tang Jin , Li Shuai , Zhao Jing , Gao Xiang 
Published: September 2020

Abstract
The potential of NK cells in cancer therapy has aroused increasing attention. While highly immune deficient mice, NCG,with null mutation on Prkdc and IL2Rγ,is ideal for human immune reconstitution, the reconstitution of human NK cells is still challenging. This is mostly because species barrier on cytokines which are necessary for the proliferation and maturation of NK cells. To this end, we established a human IL-15 (hIL-15) knock-in NCG (NCG-hIL-15), which produces hIL-15 under physiological level and pattern.NCG-hIL-15 implanted with either purified human NK cells or hematopoietic stem cells (HSCs) could give rise to a longer-term maintenance of human NK cell when compared to NCG. These long-lived NK cells retained the ability of migration, as evidenced by their wide distribution in various organs (e.g. spleen, liver, and lung). We also tested the expression profiles of killer cell immunoglobulin-like receptor (KIR) molecules, as well as the secretion of granzyme and perforin, results suggested normal activity and function of these reconstituted NK cells. Moreover, after human NK cell reconstitution, efficient tumor growth inhibition of human NK-specific agonist was shown in tumor engrafted NCG-hIL15. Taken together, NCG-hIL-15 mice is a novel mouse model for studying human NK cell biology and human NK-mediated cancer immunotherapy in vivo.
Citation Format: Cunxiang Ju, Mingkun Zhang, Dan Wu, Dingyu Wang, Hongyan Sun, Jin Tang, Shuai Li, Jing Zhao, Xiang Gao. Human interleukin 15 (IL15) humanized NCG mice support the human natural killer cells reconstitution and development [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 5613.
Abstract 5621: Humanized rat model
Authors: Adedeji Bisoye Towobola , Noto Fallon K. , Jamling Tseten Yeshi , Argawal Yash , Beatty Cole Jamison , Ho Sara Grace , Das Antu , Salunke Rajeev Kishore , Bility Moses Turkle 
Published: September 2020

Abstract
The development of Novel cancer immunotherapies has relied on the use of immune humanized mice. The field of study can be enhanced using an immune humanized rat, which will provide several advantages over the currently available humanized mouse models.
The SRG OncoRat®; A Rag2/Il2rg double-knockout rat supports the growth of larger tumors for serial fine needle biopsies to assess immune infiltration and serial blood draws for assessing human immune development and tumor biomarkers in real-time throughout an efficacy study. Using this rat, we developed a novel autologous human skin and immune cells-humanized rat model by co-engrafting full-thickness human-fetal skin and autologous fetal lymphoid organoids under the kidney capsule along with intravenous injection of autologous fetal-liver derived hematopoietic stem cells, thus termed, human skin-immune system humanized rat model (hSIS-humanized rat). We demonstrated the development of adult-like, full-thickness human skin and human lymphoid organoids along with human immune cells. Methicillin-resistant Staphylococcus aureus inoculation in the human skin results in infection and skin pathology, thus recapitulating clinical outcomes. This model will enable in vivo mechanistic studies for development and evaluation of novel therapeutics for skin infectious disease and may also provide a model for establishing skin grafts of patient-derived melanoma tumors to investigate melanoma metastasis and response to therapies. In addition, engrafting the rat with human lymphoid organs and human immune cells may provide a similar platform to the BLT mouse for immunotherapy studies.
Furthermore, we demonstrated the presence of Human CD45+, CD3+, and CD20+ cells in peripheral blood, spleen, and bone marrow of the SRG OncoRat® engrafted with human PBMCs, hence humanizing the rat's immune system. These immune humanized rat models may be beneficial for evaluating immunotherapies in human cancer models, including assessment of immune cell infiltration through fine needle biopsies.
Citation Format: Bisoye Towobola Adedeji, Fallon K. Noto, Tseten Yeshi Jamling, Yash Argawal, Cole Jamison Beatty, Sara Grace Ho, Antu Das, Rajeev Kishore Salunke, Moses Turkle Bility. Humanized rat model [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 5621.
Abstract 5689: CH7233163, a mutant-selective EGFR inhibitor, overcomes osimertinib resistant EGFR-T790M/C797S
Authors: Kashima Kenji 
Published: September 2020

Abstract
EGFR tyrosine kinase inhibitors (TKI) show dramatic improvement of treatment for EGFR-mutated lung cancer, but these clinical efficacy is limited by acquired drug resistance. Osimertinib is a clinically approved covalent EGFR-TKI and has been demonstrated to overcome the EGFR-T790M, the most relevant acquired resistance to first-generation EGFR-TKIs. However, the C797S mutation, which impairs the covalent binding between EGFR and osimertinib, induces resistance to osimertinib. Currently, there are no effective therapeutics to inhibit EGFR triple-mutation (C797S/T790M/activating-mutation). In this study, we identified CH7233163, a novel EGFR TKI, capable of overcoming the EGFR triple-mutation. CH7233163 inhibited preferentially enzymatic activity of EGFR triple-mutation, and showed potent antitumor activity against the tumor with EGFR triple-mutation in vitro and in vivo. Crystal structure of EGFR triple mutant in complex with CH7233163 shows that this compound interacted with multiple amino acids located in the ATP pocket of EGFR. This binding mode suggests that CH7233163 non-covalently but potently competes ATP without steric hindrance with T790M. Consistent with this insight, CH7233163 also showed potent antitumor activity against the tumors harboring EGFR single activating-mutation (deletion 19 or L858R) and double-mutation (T790M/activating-mutation) in vitro and in vivo. These result suggested that CH723316 may become a potential therapeutics to overcome osimertinib resistance in EGFR-mutated NSCLC.
Citation Format: Kenji Kashima. CH7233163, a mutant-selective EGFR inhibitor, overcomes osimertinib resistant EGFR-T790M/C797S [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 5689.
Abstract 569: Rational combination approaches with CRM-1 inhibitor in ovarian cancer
Authors: Handley Katelyn F. , Rodriguez-Aguayo Cristian , Ma Shaolin , Kim Mark S. , Ram Prahlad T. , Sood Anil K. 
Published: September 2020

Abstract
Background: After the frontline treatment, the majority of patients diagnosed with ovarian cancer at an advanced stage will develop recurrence (80%); thus, there is an urgent need for novel therapeutic approaches.
Purpose: Selinexor is a CRM1 inhibitor, which is a selective inhibitor of nuclear export and has demonstrated antitumor effects in ovarian cancer. Here, we examined the efficacy of selinexor alone and in combination with PARP inhibitors in ovarian cancer. As Selinexor reduces the expression of DNA damage repair proteins, we hypothesized that this combination should reduce the number of DNA damage repair proteins, resulting in synergistic tumor cell death.
Methods: Western blot was performed to assess baseline CRM1 expression in eleven ovarian cancer cell lines. We tested the efficacy of selinexor alone and in combination with olaparib in ovarian cancer cells (A2780ip2, OVCAR5). Combination Indices (CI) were calculated with CompuSyn and were considered synergistic when CI less than 1 for classic MTT arrays. We used the annexin V-FITC/PI assay to assess apoptosis as a result of selinexor and olaparib alone or in combination.
Results: Baseline CRM1 expression was positive in eight of eleven ovarian cancer cell lines tested. The average IC50 in A2780ip2 for selinexor and olaparib was 0.09uM and 0.97uM, respectively. The median combination index was 0.81 in A2780ip2 and 0.92 in OVCAR5 cells. Flow cytometry demonstrated an additive effect of selinexor and olaparib in A2780ip2 cell apoptosis (7.34% and 7.93%, respectively, v. 17.86%).
Conclusions: Selinexor demonstrates synergistic activity with olaparib, and this combination warrants further investigation.
Citation Format: Katelyn F. Handley, Cristian Rodriguez-Aguayo, Shaolin Ma, Mark S. Kim, Prahlad T. Ram, Anil K. Sood. Rational combination approaches with CRM-1 inhibitor in ovarian 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 569.
Abstract 5702: A spatially resolved single cell atlas of the human breast
Authors: Kumar Tapsi , Nguyen Quy , Nee Kevin , Bai Shanshan , Hu Min , Sei Emi , Wood Anita , Wiley Jie , Chen Hui , Contreras Alejandro , Teshome Mediget , Chen Ken , Lim Bora , Lawson Devon , Navin Nicholas , Kessenbrock Kai 
Published: September 2020

Abstract
The human breast consists of lobules connected to an intricate network of ducts that are surrounded by fatty tissues, designed to produce and transport milk to nourish offspring. Histopathology has identified 10 major cell types based on morphological features but have provided limited information on cell states - the transcriptional programs of cell types that reflect different biological functions. In this study we have generated an unbiased ‘cell atlas' of the normal human breast to define the cell types and cell states using single cell RNA sequencing and spatial transcriptomics methods. We performed 3' microdroplet based single cell and nuclei RNA sequencing of 248,687 stromal cells and 89,301 nuclei from 24 women with pathologically normal breast tissues that were collected from mastectomies and reduction mammoplasties. Unbiased expression analysis identified four major cell types: adipocytes, epithelial cells (luminal and basal), fibroblasts and endothelial cells and defined their transcriptional programs. Additionally, 8 minor cell types were identified in the breast, including macrophages, T-cells, natural killer cells, mast cells, pericytes, apocrine cells, neurons and smooth muscle cells. Our data revealed hundreds of novel markers of these cell types and defined their transcriptional programs and cell states. Most cell types had multiple transcriptional programs including luminal epithelial cells (hormone receptor positive and secretory), basal epithelial cells (myoepithelial or basal), endothelial cells (E1, E2, E3), myeloid cells, T-cells and fibroblasts (F1-F4) and provided insight into developmental lineages. We further delineated the spatial organization of these cell types and cell states within the tissue architecture using spatial transcriptomics assay. The breast cell atlas data provides an invaluable normal reference for the research community to understand how normal cell types are reprogrammed in diseases such as breast cancer.
Citation Format: Tapsi Kumar, Quy Nguyen, Kevin Nee, Shanshan Bai, Min Hu, Emi Sei, Anita Wood, Jie Wiley, Hui Chen, Alejandro Contreras, Mediget Teshome, Ken Chen, Bora Lim, Devon Lawson, Nicholas Navin, Kai Kessenbrock. A spatially resolved single cell atlas of the human breast [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 5702.
Abstract 5722: Acquired stemness by luminal cells
Authors: Karthaus Wouter , Hofree Matan , Choi Danielle , Linton Eliot L. , Turkekul Mesruh , Bejnood Alborz , Carver Brett , Gopalan Anuhandra , Laudone Vincent , Biton Moshe , Chaudhary Ojasvi , Masilionis Ignas , Mazutis Linas , Pe'er Dana , Regev Aviv , Sawyers Charles 
Published: September 2020

Abstract
Rare cell types in the prostate are reported to have stem cell properties based on organ regeneration potential following castration. Here, we use single cell RNA-seq (scRNA-Seq) to characterize these populations from the murine and human prostate in hormonally intact and androgen deprived conditions. Prostate cells from hormonally intact mice partitioned into one large subset of basal epithelial cells, another large subset of luminal epithelial cells, which we designate luminal 1 and two rare luminal populations: luminal 2 and luminal 3. Luminal cells that persist following castration display enhanced organoid regeneration potential, particularly within 1-2 days of androgen addback, and contribute equipotently to prostatic regeneration as revealed by lineage tracing. This regeneration is mediated, in part, through the orchestrated expression of Nrg2, Igf1, Fgf10 and Rspo3 by distinct populations of androgen-responsive mesenchymal and smooth muscle cells. Thus, luminal cells that persist post-castration undergo a cell state change that primes a proliferative response to microenvironment signals, analogous to other models of tissue injury such as liver damage.
Citation Format: Wouter Karthaus, Matan Hofree, Danielle Choi, Eliot L. Linton, Mesruh Turkekul, Alborz Bejnood, Brett Carver, Anuhandra Gopalan, Vincent Laudone, Moshe Biton, Ojasvi Chaudhary, Ignas Masilionis, Linas Mazutis, Dana Pe'er, Aviv Regev, Charles Sawyers. Acquired stemness by luminal cells [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 5722.
Abstract 5723: Loss of stemness and lineage commitment in the intestinal crypt
Authors: Habowski Amber , Flesher Jessica , Bates Jennifer , Ganesan Anand , Edwards Robert , Shi Yongsheng , Hertel Klemens , Waterman Marian 
Published: September 2020

Abstract
The stem cell niche in intestinal crypts maintains multi-potent stem cells that constantly replenish all cell types in the mucosal lining of normal tissue. As daughter cells exit this niche, a loss of high Wnt signaling triggers rapid differentiation and lineage commitment to secretory or absorptive trajectories. Although signaling pathways such as Wnt and Notch are known to play a role in this differentiation and commitment, the mechanism that allows for such rapid action is not well understood. To investigate this, we used a novel flow sorting protocol to sort mouse colon crypt cells and then performed deep bulk RNA-sequencing. During differentiation we uncovered a gradual loss of stem cell markers and a sharp increase in alternatively spliced and alternatively polyadenylated genes. This indicates mRNA processing may be one of the first loss-of-stemness signals in the intestine. We also were able to find lineage specific activity for the unfolded protein response and further resolution on other signaling pathways. This unique sorting procedure will aid further study of crypt dynamics and this rich dataset provides an unprecedented resolution of the intestinal crypt processes including differentiation and lineage commitment along with a baseline plasticity potential of crypt cells.
Citation Format: Amber Habowski, Jessica Flesher, Jennifer Bates, Anand Ganesan, Robert Edwards, Yongsheng Shi, Klemens Hertel, Marian Waterman. Loss of stemness and lineage commitment in the intestinal crypt [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 5723.