bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effect of mechanical cues on T cell killing of cancer cells
Authors: Dada Hannah , Do Kun , Cartagena-Rivera Alexander , Altan-Bonnet Grégoire 
Published: August 2023

Abstract
Cancer immunotherapies have focused on strategies to strengthen immune cell activation and effector function. These treatment strategies enhance the effector function of immune cells, particularly cytotoxic T cells (CTL), driving tumor eradication. Methods to improve the killing mechanisms of CTLs are not well studied and largely focused on assessing/strengthening the antigenic responses of T cells. We hypothesize that the mechanical landscape of CTL-tumor interactions can also drastically impact CTL cytotoxicity. By altering the physical context for T cell recognition of tumors, we measured cytotoxicity changes and subsequently used these changes in killing capacities to train and generate a quantitative parametrization of T cell killing. CTLs utilize cytokine expression to kill and shape the inflammatory milieu. Taking advantage of a robotic platform that enables the automatic collection of cytokines, we assembled datasets of T cell attacks of tumor cells grown on substrates with varying stiffness levels. The stiffness levels were altered using polyacrylamide (PAA) gels and ranged from 0.2kPa to approximately 113 kPa. Preliminary results from two different models (i.e., mouse and human TCR-transgenic T cells) suggest that there exists an optimal stiffness value for maximal response by T cells. These results suggest that CTLs utilize biophysical cues in addition to molecular cues (antigen) to properly elicit an immune response—modeling and fine-tuning these mechanical cues can be a novel and effective approach to improving cancer immunotherapies.
Elevated cholesterol uptake by T cells is a biomarker for the efficacy of immune checkpoint inhibitors in mouse models for triple negative breast cancer
Authors: Ciavattone Nicholas G , Guan Jenny , Farfel Alex , Desmond Tim , Stauff Jenelle , Brooks Allen , Luker Gary 
Published: August 2023

Abstract
Predicting the outcome of cancer immunotherapy can help avoid the cost and side effects of ineffective treatment and help develop effective treatment strategies. T cells take up cholesterol during activation, which is necessary for their proliferation. However, there is no literature linking the uptake of cholesterol as a biomarker for T cell activation and less so as a biomarker for immune checkpoint inhibitor therapy (ICI). Our research group has developed a novel radioactive cholesterol analog called 18F-FNP-59, which can be used to measure the uptake of cholesterol in live cells and within living organisms. We hypothesize that elevated cholesterol influx in T cells of the tumor microenvironment is linked to T cell activation and predicts the success of cancer immunotherapy in triple negative breast cancer (TNBC) models. We found that the elevated uptake of exogenous labeled cholesterol analogs functions as a biomarker for T cell activation. When comparing ICI-responsive E0771 tumors to non-responsive AT3 tumors, we found that the uptake of a fluorescent cholesterol analog was significantly higher in the T cells of the ICI-responsive tumors both in vitro and in vivo. Using the 18F-FNP-59 compound, we found that the uptake of cholesterol by T cells was further enhanced in ICI-responding tumors that received anti-PD-1 treatment. These results suggest that the uptake of exogenous cholesterol analogs by tumor-infiltrating T cells may be able to predict T cell activation and the success of ICI therapy. In future studies, we will expand the tumor models and types of cancers that we observe. Additionally we will examine the relationship between T cell cholesterol trafficking and invasion of T cells into the tumor using imaging techniques.
Investigating the Role of HDAC8 in CD4+ T Cells
Authors: Wang Jing , Yang Weiqin , Zhou Jingying , Chen Zhiwei , Cheng Alfred Sze-Lok 
Published: August 2023

Abstract
T cell exclusion restricts immune checkpoint blockade (ICB) efficacy for some hepatocellular carcinoma (HCC) patients. Epigenetic therapy provides new opportunities to rewire transcriptional programs. We have recently elucidated that pharmacological inhibition of histone deacetylase 8 (HDAC8) resulted in increased CD8+ T cell infiltration and enhanced ICB therapy efficacy in HCC. Moreover, this combined immunotherapy induced durable survival benefits even after tumor rechallenge accompanied by elevated memory T cells (Sci Transl Med, 2021). In this study, we found that HDAC8 inhibition increased memory T cells in HCC tumor-bearing mice. Interestingly, HDAC8 knockdown in tumor cells did not influence the effect of pharmacological HDAC8 inhibition in T cell activation and cytotoxic function, suggesting an immune cell-intrinsic role of HDAC8. HDAC8 was highly expressed in T cells instead of other immune cells. Notably, HDAC8 inhibition upregulated memory-related gene expression in CD4+ T cells, but not in CD8+ T cells. Besides, HDAC8 inhibition increased acetylation of structural maintenance of chromosome 3 (SMC3ac), a ring-shaped cohesin component in topologically entrapping enhancers and promoters which may modulate gene expression. Moreover, inhibiting HDAC8 increased SMC3ac binding onto the targeting gene promoter region and induced mRNA expression in CD4-expressing Jurkat cells. These data suggest that HDAC8 inhibition exerts memory T cell formation through SMC3ac-mediated epigenome reprogramming in CD4+ T cells, thus providing insights into developing effective cancer immunotherapy.
Acknowledgment: This work is supported by the RGC Collaborative Research Fund (C4045-18W), Li Ka Shing Foundation, TRS T11-706/18-N and EpiHK.
Sex-specific T cell exhaustion drives differential immune responses in glioblastoma
Authors: Lee Juyeun , Nicosia Michael , Silver Daniel , Lathia Justin 
Published: August 2023

Abstract
Sex differences in glioblastoma (GBM) incidence and outcome are well recognized; males show higher incidence and worse prognosis than females. Emerging evidence suggests that these extend to genetic/epigenetic and cellular differences, including immune responses. However, the mechanisms driving immunological sex differences are not fully understood. Using orthotopic GBM animal models, we recapitulated these sex differences in immune-competent B6 mice but not in immune-deficient mouse strains (NSG and RAG1KO). By depleting CD8+ T cells, we further confirm that CD8+ T cells play a critical role in driving sex differences in survival. Flow cytometry analysis revealed that male CD8+ T cells from tumor expressed higher levels of exhaustion markers such as PD-1, CTLA-4 and Tox, with higher frequency of progenitor exhausted T cell subset whereas female tumor contained more effector-like CD8+ T cells expressing elevated cytokine production. Anti-PD-1 antibody treatment exclusively extended the survival of male mice via inducing decreased exhausted CD8+ T cell subsets and increased effector function and proliferation. Survival analysis on bone marrow chimera model and adoptive transfer model indicated that T cell-mediated tumor control was predominantly regulated in a cell-intrinsic manner, as the sex of donor cells was a critical contributing factor. Finally, we confirmed these observations in GBM patient tumor samples with male tumors having a higher frequency of progenitor exhausted T cells with increased TOX expression. Taken together, these findings demonstrate sex-specific pre-determined behavior of T cells is critical in inducing sex differences in GBM progression and immunotherapy responses.
Supported by grants from NIH (R35 NS127083, P01 CA245705).
Radiotherapy-elicited cGAMP transfer by LRRC8 VRAC promotes anticancer T cell response
Authors: Xiao Hui , Cao Limin , Wang Li , Jia Xin-Ming 
Published: August 2023

Abstract
In addition to conducting Cl− efflux in response to cell swelling, LRRC8 VRACs have been implicated in transporting cGAMP to the bystander cells reinforcing the IFN-I response to combat HSV-1 infection. However, a plausible role of LRRC8 VRAC-mediated cGAMP transfer in cancer immunity and autoimmunity has not been explored. Here, we demonstrate that the radiation-triggered regression of implanted tumors largely depends on the host expression of Lrrc8a, especially in the T and dendritic cell compartments. Inducibly ablating Lrrc8a expression through tamoxifen or T cell-specific depletion of Lrrc8a through CD4-Cre led to diminished CD8 T cell infiltration. Moreover, the loss of Lrrc8a also impaired the expression of granzymes and IFN-γ in the tumor-infiltrated CD8 T cells. Mechanistically, our data implicate that tumor-derived cGAMP and ATP work together to trigger STING activation and IRF3/IRF7-mediated IFN-β expression in CD8 T cells. Remarkably, T cell-derived IFN-β feeds back to empower CD8 T cell’s cytolytic capacity through the induction of a subset of ISGs. In addition to radiotherapy, our observation that LRRC8 VRAC regulates anticancer immunity also extends to chemotherapy. Together, our study supports the emerging notion that both radio- and chemotherapies have the ability to boost anticancer immunity and reveal a crucial role for LRRC8 VRAC-mediated cGAMP transport in this process.
Supported by grants from National Natural Science Foundation of China.
T cell expressed microRNA-155 promotes antitumor immunity and immune checkpoint blockade responses in colon cancer through repression of Ship1
Authors: Tang William W , Ekiz H Atakan , Voth Warren P , Bauer Kaylyn M , Barba Cindy , Ghazaryan Arevik , Nelson Morgan , Lee Soh-Hyun , Weiss Allison , Thompson Jacob , Round June L , O’Connell Ryan M 
Published: August 2023

Abstract
Colorectal cancer (CRC) is the 2nd deadliest cancer globally and has the 3rd highest incidence and mortality in the United States. Advanced-stage disease is incurable by current clinical standards. Thus, alternative treatments are being explored, including immune checkpoint blockade (ICB) to prevent T cell inactivation by tumors. ICB induces a robust response in patients with microsatellite instability-high (MSI-H) metastatic CRC. However, most CRC patients are not responsive to ICB. Our analysis revealed MSI-H patients exhibit an immune cell-mediated inflammatory tumor microenvironment (TME) that is associated with the expression of microRNAs (miRs) linked to immune cell regulation. By studying T cell-associated miRs in MSI-H colon cancer patients, and through Crispr-Cas9 based in vivo screening, we identified T cell miR-155 as the miR most highly associated with ICB responsive patients. T cell miR-155 was necessary for ICB responses in multiple colon cancer preclinical models and promoted Th1-helper T cell and cytotoxic T lymphocyte enrichment in the TME, reflecting a robust immune response. T cell miR-155 controls ICB response and cancer immunity by repressing a network of target genes, most notably SHIP1. Loss of T cell miR-155 derepressed SHIP-1 gene expression in our study. Additionally, SHIP-1 was expressed in human tumor-associated T cells, with lower expression correlating to better patient outcomes. In vivo, we demonstrated that SHIP-1 inhibition partially restored tumor immunity in mice specifically lacking T cell miR-155. These findings suggest a dependence on T cell miR-155 and its targets for effective cancer immunity and ICB efficacy in colon cancer, identifying miR-155 as a candidate biomarker and therapeutic.
Supported by grants from NIH (F30 CA260977)
Identifying mechanisms driving macrophage differentiation and function in colorectal cancer
Authors: Saunders Ian T , Samakai Elsie , Brodeur Scott 
Published: August 2023

Abstract
The tumor microenvironment (TME) plays a critical role in cancer proliferation, metastasis, & responses of patients to treatment. Within the TME, tumor-initiated signaling results in aberrant activation of stromal & immune cell populations to suppress immune surveillance, induce angiogenesis, & establish premetastatic niches. Macrophages, a subset of antigen presenting myeloid cells, are the most abundant immune cell in the TME & participate in immune regulation during tumorigenesis. They functionally exhibit both anti & pro-tumorigenic properties as well as demonstrate the plasticity to repolarize between phenotypes based on environmental cues. Additionally, macrophage subset predominance within the TME & tumors have been correlated to favorable prognoses in some cancers. However, there is a lack of consensus for their role in colorectal cancer (CRC) progression. In this study, we investigated CRCs with distinct mutations, grade, & origin for their ability to drive macrophage differentiation & function in vitro. Human CRC cells were used to create multicellular tumor spheroids to better recapitulate in vivo tumor biology. CD14+ monocytes were polarized into macrophage subsets (M0/M1/M2) by cytokine/growth-factor exposure (GMCSF/MCSF/IFNγ/IL4/IL6/IL10) & co-cultured with each model. Spheroid proliferation & inflammatory profiles alone/in co-culture were assessed by live-cell microscopy, viability, & multiplex assays. Macrophage polarization, spheroid infiltration, & anti-tumor activity were examined by flow-cytometry, live-cell microscopy, & functional assessments. Overall, this study demonstrates that it may be useful to direct targeted therapies to influence macrophage polarization in CRC patients.
Supported by funding from Janssen Pharmaceutical Companies of Johnson & Johnson, Inc. under the direction of the DPDS Postdoctoral Fellowship Program and Biologics Discovery Division - Exploratory Biology Department.
Therapeutic CAR T cells differentiate into diverse memory and exhausted T cell subsets in vivo to provide long-term protection from relapse and metastasis of solid tumors
Authors: Reed Steven James , Tower Samantha J , Khanna Asheema , Kalia Vandana , Sarkar Surojit 
Published: August 2023

Abstract
Canonical memory T cell differentiation is programmed during the early stages of priming by a multitude of variables such as TCR signal strength, duration of antigenic exposure, and the milieu of cytokines. A combination of such stochastic variables in vivo, drive a heterogeneity in T cell priming that leads to the generation of distinct subsets of T cell effector and memory lineages. Contrarily, CAR T cell priming is largely homogenous and the diversity of the ensuing memory CAR T cells following adoptive immunotherapy remains largely unknown. In this study, we show that following adoptive immunotherapy into solid tumor bearing mice, syngeneic CAR T cells differentiate into distinct memory T cell subsets. In the peripheral tissues and lymphoid organs, CAR T cells subsets of central memory (TCM), effector memory (TEM) and resident memory (TRM) were fully represented. Furthermore, within the tumor microenvironment, distinct lineages of exhausted T cells such as progenitor exhausted (TPEX) and terminally exhausted (TEX) CAR T cells were found. When benchmarked against gold-standard T cells programmed in vivo by viral infections, the subsets of CAR memory and exhausted T cells demonstrated the hallmark qualities of protection from metastasis, and responsiveness to checkpoint blockade, respectively. These findings are among the first to demonstrate that CAR T cells differentiate into a full-spectrum of memory and exhausted T cell subsets in therapeutic applications against solid tumors. Comprehensive understandings of CAR T cell memory and exhaustion programs would be critical in developing therapeutic CAR T cells that provide long-term protection from tumor relapse, tumor metastasis and responsiveness to checkpoint blockade.
Research reported in this publication was supported by the National Cancer Institute of the National Institutes of Health under Award Number F32CA265056. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health
Gm2a as a novel regulator of CD8+ T cell threshold of activation in transplantation
Authors: Bazzano Julia Miranda R , Baecher Kirsten , Liu Danya , Tong Katherine , Fribourg Miguel L , Cravedi Paolo , Ford Mandy L 
Published: August 2023

Abstract
Transplant patients must remain on toxic immunosuppression for life, underscoring the need to identify immunotherapeutic targets that may facilitate transplant tolerance. We recently showed that kidney transplant recipients that remained stable following withdrawal of mainstay immunosuppression exhibited increased levels of the glycosphingolipid-catabolizing protein Gm2a, specifically in CD8+ T cells, compared to patients who went on to reject their allografts. While the role of Gm2a in lysosomal glycosphingolipid degradation in neurons is well known, little is known about Gm2a function in the immune system. To investigate this, we performed skin graft surgery in WT vs Gm2a−/− mice. Results show that Gm2a deficiency significantly increased allograft rejection relative to WT counterparts. Moreover, adoptive transfer of donor-reactive WT vs. Gm2a−/− CD8+ T cells into WT hosts resulted in increased accumulation of donor-reactive T cells and accelerated allograft rejection in recipients of Gm2a−/− CD8+ T cells as compared to recipients of WT CD8+ T cells. This increased accumulation was likely due to increased proliferation, as in vitro studies revealed enhanced proliferation of CTV-labeled Gm2a−/− vs. WT CD8+ T cells. Interestingly, Gm2a−/− CD8+ T cells exhibited increased Kb/SIINFEKL tetramer staining and sustained TCR expression following antigen stimulation compared to WT CD8+ T cells. Finally, Gm2a−/− CD8+ T cells exhibited increased responsiveness to low dose and low-affinity peptide compared to WT cells. In conclusion, these results identify Gm2a as a novel, CD8+ T cell-intrinsic regulator of the T cell activation threshold that directly impacts CD8+ T cell-mediated allograft rejection.
Supported by grants from NIH (R01AI164716)
Variegated levels of alloreactive T cell dysfunction in transplantation tolerance determine graft vulnerability to infection-triggered rejection
Authors: Allocco Jennifer Brianne , McIntosh Christine , Wang Peter , McKeague Michelle , Wang Ying , Cassano Alexandra , Xie Stephen Z , Mora-Cartin Ricardo , Chen Luqiu , Sattar Hussain , Yin Dengpin , Zhang Zheng J , Chong Anita S F , Alegre Maria-Luisa 
Published: August 2023

Abstract
While advances have been made in therapies that achieve tolerance in mouse models of transplantation, successful tolerance remains vulnerable to inflammatory insults, which can trigger graft rejection. To identify strategies for improving the robustness of tolerance, we aim to better understand the mechanisms by which grafts are rejected after donor-specific tolerance is established. In a mouse model of cardiac allograft tolerance in which late infection with Listeria monocytogenes (Lm) can trigger graft rejection, we previously showed that host CD4+ T cells reactive to host MHC-II presenting a donor MHC-I-derived peptide (TCR75 cells) became intrinsically dysfunctional if the alloantigen persisted for 3 weeks or greater. Intriguingly, infection-dependent transplant rejection was not associated with transcriptional or functional reinvigoration of these cells. We hypothesized that there might be heterogeneity in the level of dysfunction of alloreactive T cells depending on duration of their cognate alloantigen persistence. Unlike TCR75 cells, CD4+ T cells specific for host MHC-II presenting a peptide derived from donor MHC Class II (TEa cells), a graft antigen that declines post-transplantation, retained functionality during tolerance induction. Consequently, TEa but not TCR75 cells expanded following Lm-infection, supporting a role for TEa-like cells in driving infection-triggered rejection. Prolonging T cell exposure to cognate alloantigens aggravated TEa dysfunction and rendered grafts resistant to Lm-dependent rejection. These findings demonstrate that inducing dysfunction in a broader array of allospecific T cells, by prolonging exposure to more alloantigens, lessens graft vulnerability to infections.
Supported by the UChicago Growth Development and Disabilities Training Program (T32 HD007009). AHA predoctoral fellowships (20PRE35210946, 3PRE14550022 and 15PRE22180007). NIH T32-AI007090. UChicago Cardiovascular Pathophysiology and Biochemistry Training Grant (T32 HL07237). HHMI Med-into-Grad Program training grant (56006772). NIAID Grant P01AI-97113.
Exploiting Fas-mediated autocrine apoptosis of donor alloreactive T cells as an effective approach to prevent acute graft-versus-host disease in a humanized mouse model
Authors: Yolcu Esma S , Shrestha Pradeep , Turan Ali , Shirwan Haval 
Published: August 2023

Abstract
Acute GVHD initiated by donor T cells is a barrier to allogeneic bone marrow transplantation. Pan T cell elimination from the graft is associated with failure of stem cell engraftment and delayed immune reconstitution. T cells upregulate the Fas receptor on their surface following activation and become susceptible to FasL-mediated apoptosis. The objective of this study was to target Fas-mediated apoptosis for the preferential elimination of alloreactive T cells.
We previously reported a novel form of FasL protein chimeric with streptavidin (SA-FasL). Mouse and human lymphocytes were surface modified with biotin, followed by engineering with the SA-FasL protein taking advantage of biotin’s high affinity to SA. SA-FasL-engineered cells were assessed for autocrine apoptosis of alloreactive T cells in vitro and in vivo. NSG mice transplanted with 107 human PBMC were used as an acute GVHD xenogeneic model.
SA-FasL induced autocrine apoptosis in alloreactive T cells in vitro and in vivo. Mice transplanted with SA-FasL-engineered PBMC had significantly lower numbers of human T cells in the spleen and liver analyzed five days post-transplantation compared to the recipients of SA-engineered cells. Recipients of SA-engineered PBMC developed acute GVHD with a median survival time of 15 days. In marked contrast, sixty percent of SA-FasL-engineered PBMCs recipients survived for a 60-day observation period and showed minimal clinical GVHD scores.
Transient display of SA-FasL protein on the surface of donor cells provides a practical and clinically applicable effective approach to prevent acute GVHD with significant translational potential.