bluuebunny/crossref_metadata_2025_split huggingface dataset:
HLA Class I Components Increase Pancreatic Cancer Proliferation and Migration
Authors: Sliker Bailee H , Awaji Mohammad , Nguyen Thanh , Pandey Poomy , Solheim Joyce C 
Published: January 2023

Abstract
Recent studies in transplantation models have implicated HLA class I molecules as important factors in activating downstream pathways that lead to cell proliferation and migration. In a few cancers, β2m alone has been proposed to be high in expression and to increase proliferation and migration. No research to date has reported if β2m and/or the HLA class I heavy chain promote the migration and proliferation of pancreatic cancer cells and, if so, whether β2m and the HLA class I heavy chain work together or as separate entities within the signaling pathways to promote these processes.
Using pancreatic cancer cells, we demonstrate that siRNA knockdown of β2m causes a decrease in cell migration and proliferation without altering viability. This decrease of β2m also leads to a reduction in the expression of the HLA class I heavy chain as well as downstream signaling molecules, pCREB and pFAK. Downregulation of HLA-A leads to decreased pCREB while HLA-B knockdown does not and neither heavy chain isotype has an effect on pFAK expression.
Overall, this study demonstrates the importance of β2m in cancer cell migration and proliferation through signaling pathways involving pCREB and pFAK. We show that HLA class I heavy chains are expressed by pancreatic cancer cells and that HLA-A heavy chains may be more influential in cell migration and proliferation than HLA-B heavy chains. Loss of β2m expression causes a decrease in HLA class I heavy chains, suggesting that β2m may exert some of its effects on growth and migration via the heavy chain. However, the fact that some downstream molecules are altered in cells with β2m reduction, yet no change is observed with the downregulation of either HLA heavy chain isoform, indicates that each may also work in divergent pathways.
Cross-talk between NKT cells and tumor associated macrophages in the tumor microenvironment
Authors: Courtney Amy N , Tian Gengwen , Liu Daofeng , Marinova Ekaterina , Heczey Andras , Xu Xin , Guo Linjie , Gao Xiuhua , Metelitsa Leonid S 
Published: January 2023

Abstract
CD1d-reactive Va24-invariant NKT cells (NKTs) control tumor growth via yet poorly understood interactions with tumor-associated macrophages (TAMs) and other myeloid cells. TAMs comprise M1- and M2-like subsets, but only CD163high M2-like TAMs are associated with poor outcome in neuroblastoma (NB) and other tumors. Here, we demonstrate that NKTs selectively target M2-like TAMs via contact-dependent and independent mechanisms. Upon direct contact with antigen-pulsed M1 or M2 macrophages, NKTs selectively killed the latter. Although the killing was strictly CD1d-dependent, CD1d expression between macrophage subsets was equivalent. In contrast, only M2-polarized macrophages expressed CD204, a scavenger receptor responsible for cellular uptake of CD1d ligands. Anti-CD204 blocking mAb inhibited NKT-cell cytotoxicity against M2 in vitro while lentiviral transduction with CD204 cDNA rendered M1 sensitive to NKT cytotoxicity. We also found that antigen-activated NKTs could reprogram M2- into functional M1-like macrophages via GM-CSF production. Furthermore, adoptive transfer of human NKTs resulted in a M1-like polarization of TAMs in metastatic NB xenografts in humanized NSG mice. However, tumors progressed even after NKT cell treatment, suggesting a tumor escape mechanism. Indeed, we found that exposure to activated NKTs resulted in rapid up-regulation of PD-L1 on both M2 and M1 subsets, which correlated with suppressed NKT proliferation and cytokine production. Thus, our results reveal a novel mechanism of immune regulation, in which NKTs control tumor-supportive inflammation via killing or reprogramming of M2-like TAMs. However NKT function is negatively regulated by TAMs at least in part in a PD-1-dependent manner.
Characterizing the anti-tumor response of T cell receptor gene-modified invariant natural killer T cells
Authors: Fleming-Trujillo Erica Linn , Nagato Kaoru , Scurti Gina , Nishimura Michael I 
Published: January 2023

Abstract
Human invariant NKT cells represent a distinct lymphocyte population that co-express an invariant antigen receptor and NK surface markers. Classic NKT cells express a restricted T cell receptor that recognizes lipids, such as α-Galactosylceramide (α-GalCer), in the context of CD1d. Activated iNKT cells produce a high level of cytokines, such as IFN- γ, thereby bridging innate and acquired immunity through the activation of other effector cells including dendritic cells (DCs), CD8 T cells, and NK cells. One major hurdle to studying the anti-tumor activity of iNKT cells is that they do not recognize antigens presented by classical MHC molecules and they have unknown tumor antigen specificity. Therefore, we expressed the MHC class I-restricted TIL 1383I TCR in human NKT cells to characterize their biology and tumor antigen-specific response. The 1383I TCR is a CD8 independent, high affinity TCR that recognizes the melanoma antigen, tyrosinase, presented by HLA-A2. Here, we report the expression and function of TIL 1383I TCR transduced human iNKT cells. iNKT cells stimulated with α-GalCer- pulsed DCs are transduced with the retroviral vector encoding the TIL1383I TCR. After confirming TIL 1383I TCR expression in iNKT cells by flow cytometry, we assess their anti-tumor activity and polyfunctionality by intracellular cytokine staining. TIL 1383I TCR transduced iNKT cells produced IFN-γ in response to tyrosinase loaded T2 cells and HLA-A2+/tyrosinase+ melanoma. These findings indicate that genetically engineered iNKT cells are useful to study their biology in the context of antigen specificity, as well as to generate new effector populations for the treatment of cancer patients.
Loss of humoral immunity in a mouse model of ovarian cancer
Authors: Riggs James Edward , Torres Gretel , Rodriguez Wilenny , DePierri Kelley , Goldman Naomi 
Published: January 2023

Abstract
Although a low incidence cancer, ovarian carcinoma (OvCa) has a high mortality rate due to late detection. To study the humoral immune response to OvCa we transplant ID8 cells (mouse epithelial carcinoma) into the murine peritoneal cavity (PerC). We found that as the OvCa developed PerC B cells, particularly B-1 B cells, were depleted. To assess the systemic impact of this depletion we assessed humoral immunity. We found a reduction in “natural” IgM and IgG3 production and increased IgA and IgG1. Following immunization, there was loss of the TI-2 response (FITC-dextran, FITC-Ficoll); the TI-1 (FITC-LPS) response was intact until very late stage disease. Since B-1 cells also serve a housekeeping role in apoptotic corpse clearance, we used a FACS assay and a cell-based ELISA to monitor antibodies directed at apoptotic ID8 cells. Mice with OvCa had lower titers of sera that bound apoptotic ID8 cells. Collectively these data validate the systemic loss of B-1 B cells in mice with OvCa. These results could serve to inform strategies designed for early detection of OvCa.
Redirected Adoptive Cell Transfer (ReACT)- A Multi-pronged Strategy to Treat Solid Tumors
Authors: Cui Weiguo , Xin Gang , Schauder David 
Published: January 2023

Abstract
Adoptive cell transfer (ACT) of genetically engineered T cells provides unprecedented opportunities to treat hematological malignancies. However, due to insufficient migration and anti-tumor function of transferred T cells, especially inside the immunosuppressive tumor microenvironment (TME), the efficacy of ACT is much curtailed in treating solid tumors. Pathogen-based cancer vaccines can break immunosuppression in TME, but often fail to mobilize tumor-specific T cell response with ample magnitude to eradicate established tumors. To overcome these challenges, we genetically engineered tumor-specific CD8 T cells in vitro with a second T cell receptor (TCR) that recognizes a bacterial antigen. We then transfer these dual-specific T cells in combination with intratumoral bacteria injection in an approach called redirected-ACT (ReACT) to treat melanoma in mice. The dual-specific CD8 T cells expand robustly in response to bacteria, migrate to the very site of infection/tumor, and confer tumor eradication in the majority of tumor-bearing mice. The mice cured from ReACT also develop immunological memory against subsequent tumor rechallenge. Additionally, we found that this combined approach reverts the immunosuppressive TME and recruits redirected CD8 T cells with enhanced killing ability to the very site of tumor. Overall, we provide the proof-of-principle of a promising strategy to treat various malignancies.
T cell therapy targeting oncogenic antigens in ovarian cancer
Authors: Anderson Kristin G , Bates Breanna M. , Chiu Edison Y. , Greenberg Philip D. 
Published: January 2023

Abstract
Over 20,000 women are diagnosed with ovarian cancer annually, and more than half will die within 5 years. This rate has changed very little in the last 20 years, highlighting the need for innovative therapies. Reprogramming immune system cells to target and kill ovarian cancer cells represents a potentially promising new treatment strategy. Immune T cells have the potential to control tumor growth without toxicity to healthy tissues if engineered to target proteins uniquely overexpressed in tumors. Recent technological advances have helped identify and validate Wilms’ Tumor Antigen 1 (WT1) and mesothelin (MSLN) as promising ovarian cancer antigen targets; these proteins contribute to malignant and invasive phenotypes and have limited expression in healthy cells. Using patient-derived cell lines and the murine ID8 tumor, we show that T cells engineered to express a high-affinity T cell receptor (TCR) targeting either WT1 or MSLN can kill human and murine ovarian tumor cells in vitro. Moreover, in an in vivo model of established disease with disseminated ID8 tumor, adoptively transferred TCR-engineered T cells infiltrated and preferentially accumulated within ovarian tumors, delayed tumor growth and prolonged survival. However, our data revealed that the tumor microenvironment (TME) limits the survival and killing capacity of these engineered T cells. Cellular and molecular analyses of human tumor specimens have revealed human therapy will face similar obstacles. Therefore, ongoing studies will be discussed that are further exploring strategies to overcome these common elements of the immunosuppressive TME, including direct modulation of the environment and T cell engineering to promote T cell survival and function.
Expanding the scope of WT1- and cyclin A1-specific TCR gene therapy for AML and other cancers
Authors: Perret Rachel , Valliant-Saunders Karine , Cao Jennifer W , Greenberg Philip D 
Published: January 2023

Abstract
TCR gene therapy hastens the reproducible generation of tumor-specific T cells from patients, creating a promising avenue for treating hematological and solid tumors. Several hurdles remain to making this a broadly effective modality, including variable target antigen expression, risk of tumor antigenic escape, and current therapies limited almost exclusively to HLA-A0201+ patients. We previously characterized WT1 and cyclin A1 as favorable T cell immunotherapy targets, based on high expression in malignant versus normal cells and roles in oncogenesis, reducing likelihood of loss variants. We currently have a high-affinity HLA-A0201/WT1-specific TCR in Phase I trials for the treatment of AML and NSCLC, and are now focused on developing WT1- and cyclin A1-specific TCRs for ~7 common HLA types, aiming to cover &gt;90% of patients. T cell lines were generated by stimulating donor PBMC with overlapping peptide libraries to identify new reactive epitopes, and the highest affinity T cell clones for each HLA/antigen combination identified by tetramer binding strength. TCRs were then cloned into lentiviral vectors for expression in CD8 T cells. We are currently in the process of selecting the best HLA-A0201/cyclin A1-specific TCR with which to proceed to clinical development. We are also continuing to optimize our epitope discovery protocol for the improved selection of TCRs for other HLA alleles. Safety and efficacy of selected TCRs are rigorously tested using bioinformatic screens, in vitro assays, and humanized MHC-I mouse models. Our goal is to create a molecular toolbox of off-the-shelf TCR reagents to facilitate rapid treatment of patients, using T cell immunotherapies targeted to their particular tumor antigenic profile and HLA type.
VSV immunotherapy to improve treatment outcomes in a mouse model of metastatic melanoma
Authors: Shim Kevin G. , Thompson Jill , Rajani Karishma , Kottke Tim , Vile Richard 
Published: January 2023

Abstract
Vesicular Stomatitis Virus (VSV) engineered to express tumor-associated antigens (TAA) is a promising systemic therapy against widely disseminated malignancy. Using a mouse model of metastatic melanoma, we have demonstrated that VSV immunotherapy is a rational addition to current treatment strategies. Combining VSV immunotherapy with adoptive T-cell transfer and localized radiation therapy enhances the overall survival and control of disease in mice challenged with disseminated melanoma. Furthermore, our previous data have shown that the use of stereotactic hypofractionated radiotherapy provides control of subcutaneous localized disease while systemically delivered VSV immunotherapies contribute to the control of metastatic disease. From these experiments we have demonstrated that this therapy is mediated by multiple immune subsets including CD4+, CD8+, and NK1.1+ populations. Thus, we hypothesized that the additional combination of the above clinically relevant therapies with adoptive T-cell transfer and immune checkpoint inhibitors would result in distinct and quantifiable improvements in immunological tumor control. Forthcoming data will characterize the expression dynamics of the inhibitory immune receptors PD-1 and TIM3 on effector and naïve populations (as defined by CD44 and CD62L expression) in response to each of the above therapies in order to design rationale treatment strategies against metastatic melanoma.
N-acetylcysteine protects melanoma specific cytotoxic T cells from TCR restimulation induced DNA damage and enhances tumor control in vitro and in vivo.
Authors: Scheffel Matthew J , Scurti Gina , Simms Pat , Nelson Michelle , Garrett-Mayer Elizabeth , Mehrotra Shikhar , Paulos Chrystal M , Nishimura Michael I , Voelkel-Johnson Christina 
Published: January 2023

Abstract
Clinical trials utilizing the adoptive transfer of autologous tumor antigen specific lymphocytes have demonstrated superior clinical efficacy compared to standard-of-care. However, the majority of patients do not achieve a durable complete response requiring further optimization of the therapy. The rapid expansion of therapeutic T cells increases susceptibility to Activation Induced Cell Death (AICD) following TCR restimulation in vitro. Our investigations demonstrate that TCR restimulation enhances phosphorylation of γH2AX and pSMC-1, which are markers of double-strand DNA breaks. Onset of DNA damage was accompanied by the autophosphorylation of ATM, activation of the cell-death regulator p53 via phosphorylation on Ser15, and its accumulation in the nucleus. Directed inhibition of ATM with Caffeine or KU55933 protected cells from AICD. The anti-oxidant N-acetylcysteine (NAC) prevented the onset of DNA damage and subsequent ATM/p53 activation as well as cell death. Expansion of TIL1383I transduced human T cells in NAC reduced levels of γH2AX, PD-1 and CD57 while enhancing killing of MEL624 melanoma cells in vitro. In the Pmel model, expansion of T cells in NAC increased their persistence in blood and spleen and decreased γH2AX expression. NAC treatment also increased the presence of Pmel cells in the tumor, reduced tumor growth and significantly increased survival of B16-F10 challenged mice. Taken together, these data suggest the addition of NAC to current therapeutic T cell expansion protocols has the potential to improve the quality and therapeutic efficacy of the cell product infused into patients.
Comparative genome-wide screens reveal novel regulators of Cytotoxic Lymphocyte-mediated cell death.
Authors: Menasche Bridget L , Davis Eric , Sheppard Jacob , Shen Jingshi 
Published: January 2023

Abstract
Cytotoxic Lymphocytes (CLs) are key players in the effector branch of the immune response, detecting and eliminating transformed and pathogen-infected cells. While the physiological roles of CL-mediated apoptosis are well established, many regulators of these apoptotic pathways in target cells remain poorly understood. We performed comparative genome-wide haploid genetic screens in a human cell culture model for CL killing and resistance in order to identify positive and negative regulators of target cell survival during CL-mediated killing. These regulators include genes encoding glycosylphosphatidylinositol (GPI) anchor synthesis enzymes and immune surveillance receptors. In order to identify further potential components of these pathways, we performed an additional resistance screen in a GPI-anchored protein deficient haploid cell line. We identified both common and disparate regulators between unaltered target cells and GPI-anchored protein deficient target cells. These disparate regulators underline the importance of GPI-anchored proteins in CL target recognition and killing. Some gene knockouts enriched by CL-mediated killing in both screens are also mutated in cancer. Further study of the regulators identified by these screens will advance our understanding of how the many killing pathways used by CLs shape the interaction between the immune system and cancer.
A novel threonine protein kinase regulates prohibitin 2 activity and T cell function
Authors: Robles Elisa , Padilla Diana L. , Kirken Robert A. 
Published: January 2023

Abstract
Hispanic children have the highest rate of leukemia, which causes more deaths than any other cancer among children in the United States. Acute lymphoblastic leukemia (ALL) is the most common type of childhood cancer which is characterized by rapid uncontrolled accumulation of immature lymphocytes in the bone marrow and peripheral blood. Our studies have shown that the Prohibitin (Phb) family of proteins, Phb1 and Phb2, were upregulated during normal T-cell activation and likely function to maintain mitochondrial homeostasis and survival. In this work we hypothesized that a novel threonine protein kinase regulates prohibitin 2 (Phb2) activity and T-cell function. To determine the phosphorylation status of prohibitin proteins, phosphoamino acid analysis and mass spectrometry was performed followed by the generation of unique phospho-specific antibodies to characterize the putative regulatory pathways. A novel IL-2 inducible phospho-site in Phb2 was identified and characterized in T-cells. We propose that this novel threonine protein kinase participates in a previously unrecognized IL-2 signaling pathway that regulates T-cell activation and differentiation. The data also suggest that prohibitins are differentially expressed in lymphoid tumor cell lines and may serve as novel cancer biomarkers and possible molecular targets for therapeutic intervention in hematologic malignancies.
Tumor infiltrating T cells have abnormal lipid metabolism that can be modulated by PD-L1 blockade.
Authors: Yin Yiming , Metzger Todd , Bailey-Bucktrout Samantha 
Published: January 2023

Abstract
T cell activation and function are tightly regulated by nutrient availability and cellular metabolism. The solid tumor environment has several attributes that affect the metabolism of tumor infiltrating T cells (TILs). For example, tumor cells compete with T cells for glucose and thereby limit T cell effector function. Moreover, highly hypoxic conditions that are found in the tumor environment impair the oxidative metabolism of T cells. Checkpoint inhibitory and co-stimulatory pathways have been shown to regulate T cell metabolism, indicating that cancer immunotherapies may modulate T cell function via metabolic processes. Here we show that tumor infiltrating T cells have abnormally high fatty acid uptake and lipid content, which is associated with increased expression of CD36, a fatty acid transporter. The abnormal metabolism is associated with the expression of exhaustion markers programmed death-1 (PD-1) and T-cell immunoglobulin and mucin-domain containing-3 (TIM3) and is due to the cancer microenvironment rather than antigen exposure. Blocking PD-1 signaling in vivo significantly reduced tumor progression and normalized lipid metabolism. In summary, immunometabolism including lipid metabolism can serve as biomarkers of T cell anti-tumor efficacy. Targeting immunometabolism represents a promising venue in cancer immunotherapy.
Coupling metabolic activities to regulate CD8+ T cell differentiation and fate
Authors: Shrikant Protul A 
Published: January 2023

Abstract
Metabolism plays a central role in regulating CD8+ T cell differentiation and clonal expansion. Adoptive cell therapy (ACT) of cancer requires ex vivo generation of large pools of antigen specific effector CD8+ T cells. Since, differentiated effector CD8+ T cells have short life-span, exploiting mechanisms that regulate differentiation and survival can facilitate generation of large pools of effector CD8+ T cells for ACT. Integration of cell extrinsic signals during antigen stimulation (antigen, B7.1 and IL-12) induces mTORC1 activity to regulate transcriptional programs that determine functional outcomes. Our results indicate that antigen stimulation induced mTORC1 induces the mitochondria expressed uncoupling protein 2 (UCP2) to dampen glycolysis and OXPHOS to restrict effector differentiation of CD8+ T cells. Moreover, UCP2 reduces mitochondrial reactive oxygen species (ROS) production to promote clonal expansion by restricting activation induced cell death (AICD). By identifying a regulatory role of UCP2 in metabolic programming of CD8+ T cell differentiation and fate, our studies suggest targeting UCP2 expression and activity for the ex vivo generation of effector CD8+ T cells for ACT of cancer.