bluuebunny/crossref_metadata_2025_split huggingface dataset:
Hydrogen sulfide signaling in promoting the anti-tumor T cell response
Authors: Oberholtzer Nathaniel , Mehrotra Shikhar 
Published: January 2023

Abstract
Adoptive cell therapy (ACT), a form of immunotherapy, continues to emerge as a novel therapeutic strategy for treating advanced malignancies. New approaches to improve ACT protocols are needed to enhance in vivo persistence of adoptively transferred tumor antigen-specific T cells and overcome tumor-induced immunosuppression and oxidative stress, which remain significant barriers to the clinical success of ACT. Here, we show that exogenous hydrogen sulfide (H2S), an important gaseous signaling molecule, promoted a central memory phenotype with enhanced antioxidant capacity in tumor antigen-specific T cells. In vitro, T cells treated with exogenous H2S upregulated key antioxidant enzymes and molecules associated with stemness while simultaneously downregulating immune checkpoint molecules. H2S-treated T cells also displayed an enhanced reactivity to tumor antigen, with an increase in production of pro-inflammatory cytokines as well as an increase in overall protein translation. H2S-treated T cells also displayed an enhanced ability to combat oxidative stress, with a reduced accumulation of reactive oxygen species and protection against hydrogen peroxide-induced cell death. In an in vivo murine model of melanoma, tumor-reactive T cells conditioned ex vivo with exogenous H2S demonstrated superior tumor control upon ACT. Even greater tumor control was observed when ACT-treated mice were injected systemically with an H2S donor compound. These data provide evidence that anti-tumor T cells exposed to H2S possess an enhanced metabolic and stem-like phenotype that allows them to persist in the immunosuppressive tumor microenvironment and exert greater tumor control upon adoptive transfer into tumor-bearing hosts.
Supported by NIH R01 CA236379
Targeting LSD1 rescues MHC class I antigen presentation and promotes immune checkpoint blockade response in small cell lung cancer
Authors: Nguyen Evelyn M , Taniguchi Hirokazu , Chow Andrew , Sen Triparna , Rudin Charles M 
Published: January 2023

Abstract

Purpose
Small cell lung cancer (SCLC) is an aggressive tumor with modest clinical response to immune checkpoint blockade (ICB). Repression of major histocompatibility class I (MHC-I) molecules represents a key mechanism driving ICB resistance in SCLC. Recent studies have kindled interests in the lysine-specific demethylase 1 (LSD1) as an immunomodulatory target. Our study investigated the potential functions of LSD1 in regulating MHC-I antigen presentation in SCLC.

Method
We employed the inhibitor ORY-1001 and RNA interference to assess changes in MHC-I expression in SCLC cell lines by flow cytometry. We then performed RNA-seq to characterize whole transcriptomic changes in SCLC cells following LSD1 inhibition. To explore effects of targeting LSD1 on T cell cytolysis, we co-cultured SCLCs engineered to present endogenous peptides with pre-activated cognate CD8+ T cells. Finally, we treated immunocompetent mice bearing syngeneic SCLC tumors with ORY-1001 and/or anti-PD-L1 to evaluate tumor growth and characterize intratumor immune activities.

Result
We first demonstrated that targeting LSD1 restores MHC-I cell surface expression, transcriptionally activates APP-regulatory genes, and activates tumor intrinsic immunity. We further show that LSD1 inhibition of SCLC cells enables for efficient MHC-I-restricted T cell-mediated cytolysis. Finally, the addition of LSD1 inhibitor to anti-PD-L1 therapy significantly augments intratumor CD8+ T cell infiltrates and sensitizes refractory SCLC model to ICB response.

Conclusion
Our data define a potent regulatory role of LSD1 in MHC-I antigen presentation and provide support for targeting LSD1 to overcome immune evasion in SCLC.
Supported by NIH NCI R01 CA197936 (CMR); U24 CA213274 (CMR); the Druckenmiller Center for Lung Cancer Research (CMR, TS); NIH/NCI R01 CA258784 (TS); Congressionally Directed Medical Research Programs (DOD-IITRA) LC190161 (TS); Parker Institute for Cancer Immunotherapy grant (TS); LCFA-BMS/ILC Foundation Young Investigator Research Awards in Translational Immuno-oncology (TS); the Geoffrey Beene Foundation (EMN); The Yasuda Medical Foundation (HT); NIH K08 CA-248723 (AC); and the Van Andel Institute – Stand Up to Cancer Epigenetics Dream Team grant (CMR). Stand Up to Cancer is a division of the Entertainment Industry Foundation. Research grants are administered by the American Association for Cancer Research, the Scientific Partner of SU2C. We also acknowledge the use of the MSKCC Flow Cytometry Core Facility, the MSKCC Antitumor Assessment Core, and the Integrated Genomics Core, which are funded by MSKCC Support Grant/Core Grant (P30 CA008748).
Dietary low-fiber promotes resistance to immune checkpoint inhibitor immunotherapy in the LSL-KrasG12D Lung Cancer Model
Authors: Li Qingsheng , Goggin Kevin Edward , Seo Jamie , Egilmez Nejat 
Published: January 2023

Abstract
Dietary fiber interaction with the gut microbiota impact gut microbiota composition and is likely to play a large role in affecting the efficacy of immune checkpoint inhibitors (ICIs) therapy. Here we show that a diet with low-fiber promoted lung tumor development and reduced tumor-bearing animal long-term survival in both prophylactic early disease and established late disease of murine LSL-KrasG12D lung cancer model, which displays low neoantigen burden and is refractory to anti-PD-1 antibody. In this model, the administration of anti-PD-1 in the presence of IL-17 neutralization resulted in tumor suppression, indicating that limited intrinsic neoantigen alone was not the primary determinant of resistance to ICIs therapy. Interestingly, dietary low-fiber led to reduced anti-tumor effect of anti-PD-1 and anti-IL-17 partially in gut microbiota-dependent manner. Fecal microbiota transplantation from mice fed low-fiber diet also enhanced lung tumor burden. An altered immune response with significantly reduced gut lamina propria RORgt+IL-17A+ Th17 cells, lung Foxp3+Treg cells, enhanced lung Th17 cells, but no alteration of lung gdTCR+ cells were found in mice fed low-fiber diet. Furthermore, a low-fiber diet led to significant changes in metabolites including a high level of serine in the plasma of animals. Mice fed normal chow with drinking water containing serine increased lung tumorigenesis. Our study demonstrates a functional interconnection between dietary fiber and spontaneous lung tumor development.
Supported by grant from NIH/NIGMS(P20-GM135004)
Combining Immunotherapies with Conventional Cancer Therapies in a Preclinical Model of Treatment-Resistant, High-Risk Neuroblastoma
Authors: Zebertavage Lauren K , Aiken Taylor , Erbe-Gurel Amy , Schopf Allison , Nielsen Megan , Katz Sydney , Rakhmilevich Alexander , Sondel Paul 
Published: January 2023

Abstract
Patients diagnosed with neuroblastoma (NBL) are segmented into categories, with ~50% considered high-risk and ~40% of those patients eventually succumbing to disease. Our group published a regimen (“CAIR”) combining adaptive and innate immunotherapies with radiotherapy (RT) to treat an immunologically cold, treatment-resistant model of NBL (9464D-GD2). Despite CAIR being our most effective treatment, it only cures ~40% of tumors and lacks clinical relevance due in part to toxicities. Here we aimed to test if the clinically-approved salvage therapy of temozolomide and irinotecan (T/I) and αGD2-based therapy replicated human trials and to test if CAIR can be reduced to improve relevance.
For chemo studies, mice were treated T/I and αGD2 immunocytokine (Hu14.18-IL2). T/I or Hu14.18-IL2 alone did not extend survival of 9464D-GD2-bearing mice (p=0.596 and p&gt;0.999). T/I and Hu14.18-IL2 together extended survival (p=0.036 and p=0.004) but none of these treatments resulted in cures. To test CAIR reductions, mice were treated with RT, αCD40, αCTLA4, CpG, and Hu14.18-IL2. The full regimen cured 36% of mice (8/22), CAIR minus αCTLA cured 43% (9/21), CAIR minus αCD40 cured 50% (8/16), and CAIR minus CpG cured 44% (7/16). CAIR minus RT or Hu14.18-IL2 cured no mice (0/8 each).
Demonstrating similarity to human NBLs that do not respond to salvage treatment of T/I and αGD2 therapy, 9464D-GD2 tumors are not cured by T/I and αGD2 immunocytokine (Hu14.18-IL2). In contrast, CAIR can cure 9464D-GD2 bearing mice, and our data indicate that this regimen can be further reduced without decreasing efficacy. A reduced regimen enhances both clinical relevance and our ability to add additional agents to further improve survival.
Fellowship support provided by the Hartwell Foundation.
Hallmark features of T cell dysfunction are established within hours after tumor antigen encounter
Authors: Rudloff Michael William , Favret Natalie , Zumbo Paul , Dündar Friederike , Betel Doron , Philip Mary 
Published: January 2023

Abstract
Tumor-specific CD8 T cells (TST) found in patients with cancer are dysfunctional and unable to halt cancer progression. TST dysfunction, also known as exhaustion, is encoded by specific epigenetic and transcriptional programs and thought to be induced by chronic antigen stimulation. Using a genetic liver cancer mouse model, we followed the fate of naïve TST in tumor-bearing hosts. Strikingly, we found that within hours after encountering tumor antigen, TST lost effector function, expressed high levels of inhibitory receptors, and displayed chromatin peak accessibility changes previously characterized in late-stage dysfunction. This rapid onset challenges the paradigm that chronic antigen/T cell receptor stimulation is required to induce dysfunction/exhaustion. Moreover, dysfunction-associated epigenetic features were established even prior to cell division, opposing the prevailing notion that cell divisions are required for epigenetic remodeling and T cell differentiation. To determine whether these early dysfunction hallmarks were “imprinted”, we reisolated TST from malignant lesions and transferred them into antigen-free hosts. While loss of effector function was imprinted within hours of tumor exposure, other dysfunctional hallmarks, including PD1 expression, required several days of tumor exposure and additional chromatin remodeling to become stably encoded. Our study reveals the rapid induction and epigenetic remodeling which underlies TST commitment to the dysfunctional fate, highlighting the need to target early activation-induced pathways to improve anti-cancer T cell immunity.
Supported by T32GM007347-43
Directing T cell alloreactivity against solid tumors through tumor antigen dependent TCR expression
Authors: Christie Karen Michelle , Zhang Shenyu , Schwarz Thomas A , Ma Zhengyu 
Published: January 2023

Abstract
A major challenge in T cell therapy against solid tumors is the heterogeneity of tumor antigen (TA) expression. Elimination of TA+ tumor cells often fails to suppress tumor growth due to the expansion of TA− tumor cells. To address this, we developed tumor-activated alloreactive T cells (TAATs) that display alloreactivities only in tumors. TAATs lack endogenous CD3ɛ expression and contain a tumor sensing mechanism that drives the expression of an exogenous copy of CD3ɛ in the presence of TA+ tumor cells. TAATs are therefore inactive in normal tissues due to retention of TCR/CD3 in the ER but become alloreactive in tumors. TAATs attack all HLA+ cells in tumors including tumor cells, stromal cells and cells that suppress immune functions. To generate TAATs, HLA-mismatched allogeneic T cells were stimulated with HLA class I+ breast cancer cells to expand alloreactive T cells. Endogenous CD3ɛ expression was then knocked out using CRISPR/Cas9. The T cells were also modified to express a Her2-specific synthetic Notch receptor that releases an intracellular domain containing a Gal4-VP64 transcription activator upon receptor engagement. In addition, an expression cassette with a CD3ɛ coding sequence driven by a promoter containing Gal4 binding sites was introduced to the T cells. When co-cultured with Her2+ cancer cells, TAATs restored TCR/CD3 expression and were able to degranulate and produce IFNγ in response to anti-CD3 stimulations. TAATs were also able to kill Her2+ cancer cells in co-cultures and kill Her2− cancer cells in the presence of Her2+ cancer cells. Compared with CAR T cells, TAATs may achieve superior tumor suppression through attacking a wider range of cells in tumors and by overcoming the immunosuppressive tumor microenvironment.
Identifying Key Differences Between Responders vs. Non-Responders to Immune Checkpoint Blockade for Metastatic Osteosarcoma (mOS)
Authors: Di Palma Michelle Pina , Appel Nicole , Lussier Danielle , Blattman Joseph N 
Published: January 2023

Abstract
Osteosarcoma is the most common childhood bone malignancy, with a sharp decline in survival rates upon metastasis. We have previously shown immune checkpoint blockade (ICB) of anti-CTLA-4/anti-PD-L1 for mice inoculated with a K7M2 metastatic osteosarcoma (mOS) cell line resulted in ~50% survival with complete tumor clearance, including recent analogous results upon replication of this treatment regimen. Differences in response rates to ICB are common among patients with the same malignancy across various cancers. Unlike inbred lab mice, human patients have diversity in genetic makeup and other factors, causing the discrepancy in ICB response to remain poorly understood. A culprit for these discrepancies is variance in circulating antibodies (Abs), which can indicate differences in disease presence, microbiome composition, or immune-modulating factors. Recent studies have characterized Ab patterns in sera by using microarrays with thousands of randomly sequenced peptides and incubating on them diluted sera. The Ab binding pattern on the microarray is termed an “immunosignature.” We have found that before inoculation with mOS or ICB, blood from mice displays distinct differences in immunosignatures between responders and non-responders, suggesting that the presence or absence of particular Abs may influence the outcome of ICB for mOS. Additionally, recent studies have discovered that ICB’s effectiveness can be associated with or even reliant on ICB recipients’ microbiome composition and that changes in microbiome composition can result in subsequent changes to circulating Abs. Here we analyze both differences in Ab composition and microbiome composition, highlighting their impact on ICB efficacy for mOS.
Funding support for this work has been provided by a grant through the Sarcoma Foundation of America (SFA). Additional funding support for this work has been provided by a grant through the Graduate and Professional Student Association (GPSA) at Arizona State University.
Improved anti-tumor T cell generation using rapidly differentiated dendritic cells
Authors: Minguy Annabelle , Trottier Jessica , Sanchez-Dardon Jaime , Bastien Jean-Philippe , Dave Vibhuti , Roy Denis Claude 
Published: January 2023

Abstract
After allogeneic stem cell transplantation, minor histocompatibility antigens (MiHAs), Wilms-Tumor 1 (WT1) antigen, and other peptides presented by host hematologic cancer (HC) cells can lead to the in vivo expansion of donor T cells crucial for the graft-versus-leukemia effect. While the specific ex vivo production of such leukemia-specific T cells represents an interesting treatment approach, the generation of these antigen-specific donor T cells represents a major challenge. We have developed a protocol enabling the infusion of monocyte derived dendritic cell (DC)-induced ex vivo expanded T cells specific for MiHAs, WT1 and other peptides that are preferentially expressed on hematologic cells for the treatment of relapsed HC patients. While it provides interesting results, our 42-day expansion protocol is costly and requires prolonged culture periods during which patient with HC could deteriorate rapidly. To accelerate T cell production, we compared DCs generated in 3 days (DC3) vs 9 days (DC9) using standard maturation cocktail and toll-like receptor agonist. DC3 showed small differences in expression of maturation/activation markers and a unique gene signature vs DC9. Importantly, the frequency of WT1-specific CD8 T cells, primed by DC3, was slightly higher than that in DC9 primed cultures. Upon restimulation, these DC3-primed-WT1-specific T cells expressed higher amounts of IFNγ, TNFα, and CD107a degranulation marker, and were cytotoxic against WT1-expressing autologous blast cells. Together, these data show that matured DC3 have a strong capacity to prime and expand functional anti-tumor antigen-specific CD8 T cells and thus offer most interesting features for ex vivo expansion of T cells for cancer immunotherapy.
IL-9/STAT3/fatty acid oxidation mediated lipid peroxidation contributes to Tc9 cell longevity and enhanced antitumor activity
Authors: Xiao Liuling , Yi Qing 
Published: January 2023

Abstract
CD8+ T cell longevity regulated by metabolic activity plays important roles in cancer immunotherapy. In vitro polarized IL-9-secreting CD8+ Tc9 cells exert greater persistence and antitumor efficacy than Tc1/CTL cells after adoptive transfer, but the underlying mechanism remains unclear. Here, we show that tumor-infiltrating Tc9 cells display significantly lower lipid peroxidation than Tc1 cells in several mouse models, which is strongly correlated with their persistence. Using RNA-sequence and functional validation, we found that Tc9 cells exhibited unique lipid metabolic programs. Tc9 cell-derived IL-9 activated STAT3, upregulated fatty acid oxidation and mitochondrial activity, and rendered Tc9 cells with reduced lipid peroxidation and resistant to tumor or reactive oxygen species (ROS) induced ferroptosis in TME. IL-9 signal deficiency, inhibiting STAT3 or fatty acid oxidation increased lipid peroxidation and ferroptosis of Tc9 cells, resulting in impaired longevity and antitumor ability. Similarly, human Tc9 cells also possessed lower lipid peroxidation than Tc1 cells and tumor-infiltrating CD8+ T cells expressed lower IL-9 and higher lipid peroxidation- and ferroptosis-related genes than circulating CD8+ T cells in melanoma patients. This study indicates that lipid peroxidation regulates Tc9 cell longevity and antitumor effects via IL-9-STAT3-fatty acid oxidation pathway and regulating T cell lipid peroxidation can be used to enhance T cell-based immunotherapy in human cancer.
This work was supported by NCI R01s CA200539 and CA239255, and Cancer Prevention & Research Institute of Texas Recruitment of Established Investigator Award (RR180044).
Optimization of expansion techniques for adoptive NK cell transfer in dogs with cancer
Authors: Razmara Aryana , Judge Sean J , Dunai Cordelia , Murphy William J , Rebhun Robert B , Kent Michael S , Canter Robert J 
Published: January 2023

Abstract
Natural killer (NK) cells can recognize heterogeneous cancer cell targets without prior sensitization, making them promising prospects for use in immunotherapy. We have completed first-in-dog feasibility clinical trials in dogs with cancer using both autologous and allogeneic NK cells expanded from peripheral blood mononuclear cells (PBMCs). Previously, CD5 depletion of PBMCs has been used to enrich for a CD5dim expressing subset prior to NK co-culture with an irradiated feeder line, but this can limit the yield of the final NK product. The purpose of this study was to compare ex vivo culture conditions using standard CD5 depletion versus unmanipulated PBMCs plus feeder line co-culture with 100 IU/mL rhIL-2 in matched healthy donors, hypothesizing that PBMCs plus feeder cells would generate an equivalent or superior NK product to CD5 depletion. Cell count, fold change, and viability data were collected for both techniques in 12 dogs across five time points up to day 14. A mixed-effects model analysis showed no statistical difference in calculated cell counts, overall fold change, and viability (p&gt;0.05 all) between PBMCs with feeders and CD5 depleted cells with feeders. PBMCs had a higher mean than CD5 depleted cells at day 14 in all three categories, reaching a peak mean of 677 million cells from 5 million PBMCs at day 0. Flow phenotype showed similar upregulation of NKp46 and Granzyme B expression. Killing assays against melanoma and osteosarcoma targets demonstrated comparable results among PBMCs plus feeders versus CD5 depleted NK cells (p&gt;0.05). Overall, these findings support the use of unmanipulated PBMCs plus feeder line coculture as an equivalent method to CD5 depletion in the expansion of canine NK cells for adoptive immunotherapy.
Supported by grants from National Institutes of Health/National Cancer Institute (U01 CA224166-01, 1R03CA252793, T32 CA251007-01)
Aggregation of IgM+, TCRβ+ and CD4+ Cells around melanomacrophage centers - evidence of an organized lymphoid structure in a teleost
Authors: Vaidyanathan Saraswathy P , Steinel Natalie C 
Published: January 2023

Abstract
The study of immunity in non-model organisms has the potential to reveal the evolved diversity of the immune system. However, such studies are hampered by the paucity of species-specific reagents. Advances in single-molecule RNA-ISH overcomes these setbacks and allows for the detection of potentially dozens of gene targets in a single tissue section. We examined the spatial expression of immune genes in the splenic tissue of a teleost, threespine stickleback, using the advanced multiplexed RNAScope Hiplex assay. Threespine stickleback, a small teleost found throughout the Northern Hemisphere, is an emerging immunologic model. It has been proposed that teleost humoral adaptive immunity initiates in the melanomacrophage center (MMC), a putative evolutionary precursor to germinal centers (GC), however this has not been directly tested. Teleosts lack GCs, yet still generate affinity matured antibodies in response to antigen challenge. We find that stickleback MMCs increase in size in response to antigen immunization but not to control injection, suggesting that MMC response is antigen driven, similar to the mammalian GC. Analysis of splenic tissue indicates that IgM mRNA expressing cells aggregate in discrete regions around stickleback MMCs. Similarly, CD4 and TCR-β expressing cells aggregate near IgM+ cells, comparable to the organization of B and T cells in mammalian GCs. These findings suggest that stickleback splenic tissues have an organized structure, focused around the MMCs, and support the hypothesis that MMCs could be the site of teleost adaptive immunity and the evolutionary precursors to GCs. This study also highlights the application of RNAScope in a non-model organism.
Supported by NIH RO1AI146168