bluuebunny/crossref_metadata_2025_split huggingface dataset:
Humanized Rag2 mice as patient-derived orthotopic mouse models for immune checkpoint blockade and conventional chemotherapeutic drug treatment for human solid tumors
Authors: Li Li , Zhang Xin , Maresh Grace , Mathew Neeha , Bhattarai Amita , Hellmers Linh , Yip Henry , Moret Ravan , Baste Lara McKean , Green Heather , Latsis Maria , Bardot Stephen , Margolin David 
Published: January 2023

Abstract
To evaluate therapeutic efficacy of immune checkpoint blockade (ICB) and conventional chemotherapy in pre-clinical models, we developed humanized patient-derived orthotopic xenograft (PDOX) models for colorectal cancer (CRC) and renal cell carcinoma (RCC).
Rag2 mice were humanized by donor peripheral blood mononuclear cells (PBMC) transfer. Luciferase-tagged CRC and RCC tumor cells were injected intra-rectally and intra renal subcapsular, respectively. Groups of mice (n=5–10) received ICB, anti-PD-1 and anti-PD-L1 antibodies (Nivolumab and Atezolizumab, 200 μg each/mouse) for 4 weeks, and conventional therapy of 5FU (200 mg/kg, for CRC) alone or in combination. Tumor growth was measured by weekly bioluminescent image (BLI) and tumor weight at necropsy. The presence of human immune cells and tumor infiltrating lymphocytes were confirmed by flow cytometry and immunohistochemistry staining.
Humanization was evidenced by &gt;45% HLA-ABC+CD45+ circulating human cells in both CRC and RCC models. CRC tumor weight showed significant reduction in group treated with combination of ICB and 5FU compared with untreated controls or 5FU monotherapy group (p&lt;0.05) using patient tumor cell CoCa302. The lung/liver metastasis detected by ex vivo BLI were reduced from 50% of mice in control group to 33% and 11%, respectively. For RCC, tumor size was reduced by &gt;30% from controls in ICB treated group using patient tumor cell KiCa118 and cell line SN12K1. The later had 20% reduction of liver metastasis.
The combination of ICB and conventional chemotherapies for CRC and RCC can be tested in our humanized PDOX mouse models. The effect of the combination therapy using MHC matched donors or patient autologous PBMC will be further investigated.
Radiation-Induced Immunity in Kidney Cancer Patients: Seq-ing Commonality Between Peripheral and Intratumoral T cell Repertoires
Authors: Chow Jacky Louis , Hoffend Nicholas , Abrams Scott I , Schwaab Thomas , Singh Anurag , Muhitch Jason Bryan 
Published: January 2023

Abstract
Radiation has long been part of the standard of care for cancer treatment. Clinical studies combining immunotherapy with radiation have shown promising response rates implicating radiotherapy as an inducer of tumor immunity. Due to the scarcity of human samples treated in situ with radiation, exploration of this phenomenon has been largely limited to murine models. While these data draw connections between intratumoral immune responses and wider effects in peripheral blood, advances in patient care are restricted by inherent differences between murine models and human disease. Therefore, to identify commonality between local intratumoral and systemic responses to radiation, we analyzed clinical trial samples from renal cell carcinoma (RCC) patients treated with stereotactic body radiation therapy (SBRT) at the primary tumor. With access to patient tumors and peripheral blood, we hypothesized that SBRT initiates local and systemic anti-tumor immunity. Pathway analysis from tumor RNAseq data showed SBRT-specific immune processes: TCR activation, increased costimulation, and chemokine signaling. Deeper examination through single cell RNA sequencing identified the sources of these signals. In parallel, analysis by T cell receptor sequencing (TCRseq) showed increased tumor T cell clonality with SBRT treatment. TCRseq of serial blood samples revealed tumor-enriched T cell clonotypes reaching peak abundance 2 weeks post-SBRT. This expansion was not sustained and contracted towards baseline levels 4 weeks after SBRT. Collectively, these results indicate a critical interval following radiation, wherein tumor-enriched T cell clones are expanded in circulation, that could be utilized to bolster responses to immunotherapy.
Harnessing the power of trained immunity using yeast-derived β-glucan to prevent pancreatic cancer
Authors: Geller Anne E , Ding Chuanlin , Guo Haixun , Yan Jun 
Published: January 2023

Abstract
Despite the remarkable success of immunotherapy, pancreatic cancer has yet to benefit. This creates a desperate need for therapies that target pancreatic cancer and utilize novel approaches to prevent and treat this lethal disease. Innate immune cells are vital in antitumor immunosurveillance. Recent studies have shown that innate immune cells possess a form of memory termed Trained Immunity (TI), where cells exposed to an initial stimulus undergo epigenetic and metabolic reprograming that causes hyper-responsiveness when exposed to a second signal. It is yet unknown whether inducing TI could protect against tumors, and specifically pancreatic tumors. Here we show that yeast-derived particulate β-glucan, a known TI inducer, results in 90% of an intraperitoneal dose trafficking to the pancreas. This leads to a robust, dose-dependent influx of innate and adaptive immune cells into the pancreas that have enhanced Th1 function. Despite the cellular influx, signs of pancreatitis are absent. In vivo and in vitro studies of β-glucan trained pancreatic macrophages show increased TNFα and IL-6 production when exposed to LPS, pancreatic tumor cells and their supernatant. This indicates that pancreatic macrophages can be trained to protect against pancreatic tumor cells. Depletion of various immune populations suggest that pancreatic macrophages are directly trained by β-glucan, which then stimulate NK and T-cells. In an orthotopic model of pancreatic cancer, β-glucan trained mice show reduced tumor burden and augmented antitumor functions of innate and adaptive cells. Together, this suggests that β-glucan can train pancreatic innate immune cells to have antitumor memory, which can serve as a novel approach to treating pancreatic cancer.
Dietary administration of alpha-tocopheryloxyacetic acid induces CD4+ T cell activation and prolongs survival in murine rhabdomyosarcoma
Authors: Capitini Christian M. , Szewc Fernanda , Kabakov Sabrina A , Rinella Sean P , Akporiaye Emmanuel , Redmond William L 
Published: January 2023

Abstract
Relapsed sarcomas remain essentially incurable for most children with cancer. Alpha-Tocopheryloxyacetic acid (αTEA) is a scalable semi-synthetic derivative of alpha-tocopherol that has shown promising activity preclinically and clinically in adult cancer, but has not yet been studied in pediatric sarcoma. We hypothesized that αTEA would mediate anti-tumor effects in rhabdomyosarcoma (RMS) by modulating the immunosuppressive milieu. Treatment of murine M3-9-M RMS in vitro with αTEA (2.5–100uM) resulted in increased apoptosis by flow cytometry (Annexin V+/7AAD+) and Incucyte analysis (Annexin V+) within 24 hours (p &lt; 0.05). Using an established orthotopic luciferase+ M3-9-M in vivo RMS model, αTEA-supplemented chow administered for 21 days significantly reduced tumor volume (p &lt; 0.001) and bioluminescence (p &lt; 0.05) by 28 days, resulting in a significant prolongation of survival compared to matched control chow (p &lt; 0.05), up to 4 weeks after the feed was stopped. Analysis of spleens from RMS mice fed αTEA-supplemented diets showed an increased percentage of Ki-67+ and IFNγ+CD4+ T cells (p &lt; 0.01) at the end of treatment as compared to recipients of the control diet. In addition, a decrease in the percentage of CD11b+Arg-1+ (p &lt; 0.01) and PD-L1+ (p &lt; 0.05) cells within the spleen were observed in αTEA-treated RMS tumor-bearing mice. RNA-Seq analysis of RMS tumors is underway to explore changes in αTEA vs. control-treated mice. These results suggest that αTEA causes direct induction of apoptosis of RMS in vitro and in vivo, leading to prolonged survival through enhanced CD4+ T cell stimulation and reduced myeloid cell-mediated immunosuppression in RMS-bearing mice. Further investigation of αTEA as an immunomodulating agent is warranted.
The tumor associated antigen LEDGFp75 interacts with oncogenic transcription factors in docetaxel resistant prostate cancer cells
Authors: Ortiz-Hernandez Greisha L , Sanchez-Hernandez Evelyn S , Casiano Carlos A. 
Published: January 2023

Abstract
Our group and others established that the stress oncoprotein LEDGF/p75 is a tumor-associated antigen (TAA) targeted by serum autoantibodies in certain patients with prostate cancer (PCa). LEDGF/p75 is also upregulated in clinical prostate tumors and contributes to docetaxel (DTX)-resistance in advanced PCa cells. The C-terminus of LEDGF/p75 contains a domain called the Integrase Binding Domain (IBD), which in T cells is responsible for tethering the HIV-integrase complex to transcriptionally active chromatin. In cancer cells, the IBD interacts with oncogenic transcription complexes to promote cell survival. However, the relevance of these LEDGF/p75 protein-protein interactions (PPIs) in PCa and chemoresistance has never been explored. This study aims to test if the interactions between LEDGF/p75, JPO2, Menin, and MLL in DTX-resistant PCa cells contribute to chemoresistance. Also, we want to target these interactions with repurposed HIV-based small molecule inhibitors (SMIs) of LEDGF/p75, which target the IBD, to abrogate this resistance. We demonstrated a significant co-upregulation of these proteins in DTX-resistant PC3-DR and DU145-DR cell lines, compared to the parental cells. Also, we observed nuclear co-localization by immunofluorescence microscopy. Using human anti-LEDGFp75 autoantibodies, we confirmed that endogenous proteins co-immunoprecipitate in the DTX-resistant PCa cell lines. Besides, we are evaluating repurposed HIV-based SMIs for their efficacy in sensitizing resistant cells to DTX. Our goals are to use anti-LEDGFp75 autoantibodies to establish its endogenous-interactome, contribution to transactivation of DTX-resistant genes, and target this interactome to overcome PCa chemoresistance.
Inflammation-induced tumor antigen expression on epithelial cells reveals the mechanism of their generation and provides a system for identification of new tumor targets
Authors: Jacqueline Camille , Lee Amanda , Frey Nolan , Minden Jonathan S , Finn Olivera J 
Published: January 2023

Abstract
Tumor-associated antigens (TAAs) are self-molecules abnormally expressed on tumor cells that elicit humoral and cellular immunity and are targets of effective cancer immunosurveillance. Surprisingly, immunity to TAAs is found in many healthy individuals with no history of cancer. We previously showed that other events in addition to cancer, such as viral and bacterial infections, can cause the expression of some TAAs and correlate with effective tumor immunosurveillance. We are now testing in mice the ability of two LCMV strains, one that causes acute (Armstrong) and one that causes chronic (Cl-13) infection, to elicit expression of and immune memory to TAAs and protect from a later tumor challenge. We observed that infection with the acute strain elicits immunity to specific TAAs expressed by two tumor cell lines that were then used in tumor challenge experiments. So far, we were able to show a protective effect of a previous Armstrong infection against our model of lymphoid tumor (EL4). We also developed a highly reproducible in vitro model of acute inflammation of primary epithelial cells and confirmed that short-term exposure to pro-inflammatory cytokines IL-1β, IL-6 and TNF-α leads to abnormal expression of three well known TAAs, MUC1, CEA and HER2/neu. These cells were found to activate macrophages, which should promote antigen uptake and presentation of these and other TAAs. We have used this system and 2D Difference Gel Electrophoresis (2D-DIGE) combined with mass spectrometry to identified so far seven new TAAs. Because these TAAs are generated first as disease-associated antigens (DAAs), they represent highly predictable shared antigens for safe and effective preventative cancer vaccines.
Single-cell systems neuroimmunology reveals a highly immunosuppressive microenvironment in human glioblastomas contacting the ventricular stem cell niche
Authors: Bartkowiak Todd , Barone Sierra , Mistry Akshitkumar M , Greenplate Allison R , Sinnaeve Justine , Leelatian Nalin , Hayes Madeline , Roe Caroline E , Mobley Bret C , Chambless Lola B , Thompson Reid C , Weaver Kyle D , Ihrie Rebecca A , Irish Jonathan M 
Published: January 2023

Abstract
Glioblastomas make up more than 60% of adult primary brain tumors and carry a 15 month overall survival despite aggressive standard-of-care therapy. Recent advances in immunotherapy offer an appealing alternative that may improve outcomes for patients with glioblastoma; however, clinical trials have proven unsuccessful due in part to a lack of predictive features that may inform responsiveness to immunotherapy. We have recently shown a strong correlation between 1) immune infiltration, 2) tumor cell phenotype, and 3) patient outcome. Further, patients whose tumors demonstrate radiographic contact with the ventricular-subventricular zone (V-SVZ) have reduced survival compared to patients whose tumors do not contact the V-SVZ. We therefore hypothesized that the V-SVZ acts as a previously unappreciated immunosuppressive microenvironment within the brain that promotes tumor growth by suppressing anti-tumor immunity. Primary human glioblastomas were disaggregated into single-cell suspensions and mass cytometry (CyTOF) measured &gt;30 parameters in thirteen immune populations infiltrating human glioblastomas. Cutting-edge machine-learning tools identified key differences in the abundance and phenotypes of T cells, B cells, NK cells, microglia, and peripheral macrophages infiltrating ventricle-contacting gliomas. Further, enriched expression of immune checkpoint receptors (PD-1, TIGIT, LAG-3, TIM3) correlated with ventricular contact and outcome. These results provide key insights into the immune microenvironment of glioblastomas and elucidate several clinically actionable immunotherapeutic targets that may be used to optimize treatment strategies for glioblastoma patients based on V-SVZ contact status.
The impact of the tumor microenvironment on granzyme production by CD8 T cells
Authors: Hay Zachary LeeZer , Kim Dale , Silva Alexa Lynn , Slansky Jill E 
Published: January 2023

Abstract
Granzymes are a class of proteases produced by CD8 T cells to mediate their cytotoxic activity against target cells. The objective of this project is to determine the mechanism through which CD8 T cells are executing their granzyme-mediated cytotoxic function and how the tumor microenvironment (TME) impacts the specific granzymes produced. By micro-array and qPCR we determined that CD8 T cells in the TME have altered expression of specific granzymes. Granzyme F had one of the most altered expressions of all genes examined in antigen-specific CD8 T cells taken from a spleen relative to those taken from a tumor. We further examined the effects of the TME on granzyme expression by sorting tumor infiltrating CD8 T cells on exhaustion markers and found a relationship between exhaustion and granzyme expression, characterized by a decrease in granzyme A and an increase in granzyme B mRNA expression. We will determine the influence different ex vivo culture conditions have on differential granzyme expression and function of CD8 T cells. The expression of different granzymes may have further implications for modulation of the TME through the form of cancer cell death induced, especially the increase in granzyme F which has been suggested to induce a more immunogenic form of cell death. Additionally, the impact of exhaustion on the cytotoxic activity of CD8 T cells from the TME may represent a means for exhausted CD8 T cells to regulate their cytotoxic activity by altering the granzymes they express.
Molecular cytometry identifies a wide range of translationally relevant markers in tumor and peripheral immune cells of lung cancer patients
Authors: Chattopadhyay Pratip K , Gao Guo-Jian , Taylor Ian , Guidry Kayla , Labbe Kristin , Almonte Christina , Nakamoto Margaret , Kin-Wong Kwok 
Published: January 2023

Abstract
Many immunotherapy drugs, used singly or in combination, are emerging. To realize precision oncology, and better design clinical trials, in-depth immune profiling is key. Many new technologies are available; the most promising simultaneously analyzes &gt;102 proteins and 400 mRNA cell-by-cell (molecular cytometry). Using this technology, we deeply profiled TIL, from freshly resected lung tissue, and PBMC collected at surgery (n=10).
Antibody staining was robust, with all canonical cell populations at expected frequency. We identified markers uniquely upregulated in TIL, including CXCR6, CD39, CD26, CD69, CD103, and RGS. We also asked what markers were uniquely enriched in PD1-TIL, in order to find other drug targets for patients who fail Nivolumab. We found many molecules upregulated in PD1-TIL, including CD326, CD98, LGALS3, TIM3, CD54, CD235ab, CXCL8, CD141, and CD117. We further identified precise combinations of these markers that inform design of combination immunotherapy.
We also characterized immune landscapes in metastatic disease vs. localized adenocarcinoma, finding drug targets and tumor-immune interactions are different across these settings. For example, T-cell activation markers are downregulated in metastatic tissue, e.g., HLADR (p = 1.5E-16), CD69 (p=2.9e-10), and CD38 (p=3.2e-16). CD103 was also highly downregulated (p=1.9E-14). Notably, in metastatic disease various myeloid proteins are elevated, including CD206 (p=0.002), CD32 (p=0.02), and CD61 (p=0.006). We also characterized the degree of immune exhaustion, using ratios of ZBED2:LGALS in cells. In summary, we demonstrate the utility of molecular cytometry for providing unique (and translationally-relevant) insight into lung cancer.
Inflammation synergistically promotes cigarette smoke carcinogen-induced lung tumorigenesis and determines immunotherapy efficacy
Authors: Di Yuanpu Peter , Liu Chia-Hsin , Chen Zhong , Chen Kong , Liao Fu-Tien , Keohavong Phouthone , Leikauf George 
Published: January 2023

Abstract
Chronic obstructive pulmonary disease (COPD) is a chronic lung inflammatory disease with an increased risk of lung cancer. Pseudomonas aeruginosa (PA) are opportunistic bacteria frequently found in patients with COPD, and associated with increased lung inflammation and acute exacerbations. The impacts of COPD-related inflammation, especially those induced by the PA-produced lipopolysaccharides (LPS), on lung tumorigenesis and the efficacy of immune checkpoint blockades remain poorly understood. In this study, we established a novel murine lung cancer model by treating mice with cigarette smoke carcinogen nitrosamine (NNK) combined with recurrent LPS exposure. LPS-mediated chronic inflammation in NNK-treated mice generates an immunosuppressive microenvironment characterized by the accumulation of myeloid-derived suppressive cells (MDSCs) and regulatory T cells, induced T-cell exhaustion, and enhanced PD-1/PD-L1 activity that resulted in synergistic increases of lung tumorigenesis than NNK treatment alone (&gt;8-fold increase in tumor numbers, P&lt;0.0001). Immunotherapy using anti-PD-1 antibody or anti-Ly6G antibody (to deplete MDSCs) alone reduced tumor growth, and in combination further enhanced the anti-tumor activity. We also identified immune gene signatures from the RNA transcript expression data of our murine cancer model and the published datasets of PD-1 blockade, which effectively predicted treatment responses and survival outcome in cohorts with immunotherapy and conventional treatments. Our results indicate that LPS-mediated chronic inflammation creates a favorable immunosuppressive microenvironment for tumor progression and correlates with the efficacy of anti-PD-1 treatment.
PD-L1 lncRNA splice promotes lung adenocarcinoma proliferation and metastasis via enhancing c-Myc activity
Authors: Liang Hongwei , Qu Shuang , Zen Ke 
Published: January 2023

Abstract
Although blockade of programmed death-ligand 1 (PD-L1) to suppress T cell immune responses shows great promise in tumor immunotherapy, the efficacy of such immune-checkpoint inhibition strategy is low for patients with solid tumors. The mechanism underlying the limited efficacy of PD-L1 inhibitors remains unclear. Here, we show that, via alternative splicing, human PD-L1 protein positive or negative lung adenocarcinoma all produce a long non-coding RNA isoform of PD-L1 (PD-L1-lnc), which promotes lung adenocarcinoma proliferation and metastasis. Analyses of various lung adenocarcinoma cells show that PD-L1-lnc is significantly upregulated by IFNg in a manner similar to PD-L1 mRNA. Both in vitro and in vivo studies demonstrate that PD-L1-lnc increases proliferation and metastasis, but decreases apoptosis of lung adenocarcinoma cells. Mechanistically, PD-L1-lnc directly binds to c-Myc and enhances c-Myc signaling downstream in lung adenocarcinoma cells. Therefore, targeting PD-L1-lnc/c-Myc axis provides a novel strategy for lung cancer therapy.
Low immunogenicity of intracranial tumors limits immune response to brain malignancies
Authors: Stasiak Katarzyna , Lindsay Robin , Engelhard Victor H. 
Published: January 2023

Abstract
The presence of CD8 T cells in tumors, including melanomas, is correlated with longer survival. Cutaneous melanoma is a highly immunogenic malignancy with high rate of dissemination into central nervous system but the brain is considered immunologically privileged due to the blood-brain barrier, which limits T cell entry. To determine if brain tumors are also poorly infiltrated by T cells we evaluated intracranial (IC) and subcutaneous (SC) B16 melanomas expressing different model antigens.
CD8 T cells were present in IC tumors 7 days post tumor implantation in B16 melanomas expressing chicken ovalbumin (B16OVA) or influenza M1 protein (B16M1), but infiltrates were only 10% of those observed in SC tumors. We also observed that expression of homing receptor ligands (HRLs) known to mediate T cell infiltration was lower on IC than SC tumor-associated vasculature. Despite the fact that HRL expression decreased over time in tumors grown in both locations, T cell infiltrates increased, suggesting that HRL are not the only factor limiting T cells entry into IC tumors.
To determine whether the IC tumor infiltrate size was limited by inefficient stimulation of effectors cells, we vaccinated mice to generate fully activated effector T cells able to recognize tumor associated antigen. Vaccination normalized the infiltrate sizes in IC and SC tumors. Interestingly, this was also dependent on expression of Ag by the tumor cells. This suggests that IC tumors are not able to efficiently elicit a robust immune response, possibly due to limitations on drainage to lymph nodes, while IC tumor vasculature does not impose inherently greater limitations on the ability of T cells to enter.