bluuebunny/crossref_metadata_2025_split huggingface dataset:
Interactions between the Environment and Genetics determines immune variation in rewilded mice
Authors: Oyesola Oyebola Oluwakemi , Downie Alexander E , Barre Ramya Smithaveni , Chen Ying-Han , Kiwanuka Kasalina N , Zaldana Kimberly , Howard Nina , Lee Soo Ching , Devlin Joseph , Mondragon Octavio Paloma , Herrmann Christin , Zhao Mingming , Koralov Sergie B , Cadwell Kenneth , Graham Andrea L , Loke P’ng 
Published: January 2023

Abstract
Immune responses to pathogens and vaccination can be varied with some individuals inducing optimal responses while others do not. The host genetic profile, environment and previous microbial experience could influence an individual’s response, but the relative contribution, and interactions of these different factors remains largely unknown. Here, using various multi-omics, ecological and single cell approaches, we show that release of genetic inbred strains of mice, 129-SL, PWK and C57/B6 mice, to a rewilded environment and exposure of these rewilded and laboratory specific pathogen free control mice to a helminth parasite, Trichuris muris allowed us to assess the contribution and interaction of host genotype and environment to the immune cell landscape in the blood and secondary lymphoid organs. Critically, we find that the environment has the greatest effect on circulating blood immune cells while the genetic profile has the greatest effect on the mesenteric lymph node. We also observed significant interactions between the host genetic profile, environment, and infection status in their contribution to immune cell composition, with most of the effect driven by the cells of the adaptive immune system. These findings provide a model for contribution and interactions between genetics, environment, and helminth infection in the inter-individual variation of immune responses.
A novel digital device-based adjuvant-like effect for influenza vaccine: A feasibility and pilot study
Authors: Hahm Jarang , Choi Jun-Pyo , Ayoub Ghazal , Kim Sae-Hoon , Kim Eu Suk , Noh Ji Yun , Song Joon Young , Chang Yoon-Seok 
Published: January 2023

Abstract
Viral infections such as influenza and COVID-19 have a huge impact on humans. Previous studies showed that some exercise could enhance the anti-viral antibody titers after vaccination. We developed a novel digital device designed to enhance host immune system against viral infection by acting like adjuvants. The digital device, SAT-008, is a mobile application based on an algorithm to regulate physical activity. To assess the feasibility of SAT-008, a randomized, open-label, and controlled study was conducted for 13 weeks (Oct 20 to Jan 21). A total of 42 healthy adults (mostly healthcare workers) aged 24 to 46 years were recruited and finally 32 subjects served for analysis. A single-dose of quadrivalent inactivated influenza vaccine was administered for each subject. The control group maintained daily life without using SAT-008, while the experimental group (Exp) used SAT-008 during the study. As a result, compared to the controls, the Exp showed a significant increase in hemagglutination-inhibition (HI) titers of antigen subtype B Yamagata lineage after 4 weeks of vaccination and antigen subtype B Victoria lineage after 12 weeks of vaccination (P &lt; 0.05), whereas the controls did not. As for type A influenza, there was no significant difference in HI titers between groups. In addition, the Exp was divided into high performers (HP) and low performers (LP) based on the adherence rate. The HP showed significantly higher HI titers of B Yamagata lineage in 12 weeks after vaccination than controls and LP (P &lt; 0.05) and of B Victoria lineage than controls (P &lt; 0.01). Our findings indicate a novel approach using digital device may play an important role to enhance host immune system to act as a vaccine adjuvant against viral diseases such as influenza.
Supported by grants from S-Alpha Therapeutics, Inc.
A feeder cell-free activation and expansion strategy to generate memory-like NK cells sufficient for off-the-shelf multi-dose adoptive cell therapy.
Authors: Wong Hing C , Dee Michael J , Shrestha Niraj , Chaturvedi Pallavi , Leclerc Giles M , Zhu Xiaoyun , Liu Bai , Kong Lin , Echeverri Christian A , You Lijing , Egan Jack O , Jiao Jin-An , Rhode Peter R , Becker-Hapak Michelle K , Berrien-Elliott Melissa M , McClain Ethan , Foster Mark , Pence Patrick , Neal Carly C , Kersting-Schadek Samantha , Fehniger Todd A 
Published: January 2023

Abstract
Adoptive cell therapy (ACT) using NK cells is a promising armament in the fight against cancer. Cytokine induced memory like (CIML) NK cells have been shown in clinical studies to have potent antitumor activity with superior in vivo persistence. Currently, the expansion of NK cells for clinical development is mainly based on feeder cells, which imposes significant regulatory hurdles and increases the costs for manufacturing. We have developed fusion proteins, HCW9201 and HCW9206 comprising of IL-15/IL-18/IL-12 and IL15/IL-7/IL-21, respectively, capable of priming memory-like differentiation and expanding CIML NK cell products without using feeder cells. This “Kick and Expand” strategy allows greater than 100x expansion of CIML NK cells from donor PBMCs in as little as 14 days without the use of exogenous feeder cells. Continued expansion can yield sufficient CIML NK cells for cryopreservation and multiple ACT infusions. The NK cells generated have bona fide memory-like properties: enhanced antitumor activity across multiple cancer cell lines, higher metabolic capacity, stable epigenetic demethylation of the IFN-γ promoter and increased persistence in NSG mice, when compared to conventional NK cells. In conclusion, this “Kick and Expand” process supports generation of abundant CIML NK cells for multiple ACT infusions and provides simpler, more regulatory friendly, off-the-shelf platform for generating NK cell products, including those with chimeric antigen receptor (CAR) constructs.
Cell membrane-anchored and tumor-targeted IL-12 (attIL12)-T cell therapy for eliminating large and heterogeneous solid tumors
Authors: Hu Jiemiao , Yang Qing , Zhang Wendong , Du Hongwei , Dao Long , Xia Xuqing , Fowlkes Natalie , Mahadeo Kris , Gorlick Richard , Dotti Gianpietro , Li Shulin 
Published: January 2023

Abstract

Background
Adoptive T-cell transfer has become an attractive therapeutic approach for hematologic malignancies but shows poor activity against large and heterogeneous solid tumors. Interleukin-12 (IL-12) potent antitumor efficacy against solid tumors, but its clinical application has been stalled because of toxicity. Here, we aimed to develop a safe approach to IL-12 T-cell therapy for eliminating large solid tumors.

Methods
We generated a cell membrane-anchored IL-12 (aIL12), a tumor-targeted IL-12 (ttIL12), and a cell membrane-anchored and tumor-targeted IL-12 (attIL12) armed T cells, CAR-T cells, and TCR-T cells with each. We compared the antitumor efficacy of these armed T cells in treating osteosarcoma PDX tumors and mouse melanoma tumors, and addressed the safety concerns of IL-12 expressing T cell therapy after intravenous infusions of the armed T cells.

Results
attIL12-T cell infusion showed remarkable antitumor efficacy in human and mouse large solid tumor models. Mechanistically, attIL12-T cells target tumor cells that express cell-surface vimentin (CSV) and accumulate in tumor sites. Such CSV-attIL12 interaction stimulates effector T cell to produce high levels of inflammatory cytokines (IFNγ, Flt3L, GM-CSF and etc.) only in tumors, which in turn triggers dendritic cell maturation for activating secondary T-cell responses and tumor antigen spreading. Both attIL12- and aIL12-T-cell transfer eliminated peripheral cytokine release and the associated toxic effects due to tumor-specific T cell distribution and compartment-dependent cytokine secretion.

Conclusions
This novel approach sheds light on the safe application of IL-12–based T-cell therapy for large and heterogeneous solid tumors.
Supported by R01 CA200574
Close proximity of immune and tumor cells underlies response to BRAF/MEK-targeted therapies in metastatic melanoma patients
Authors: Yan Chi , Chen Sheau-Chiann , Ayers Gregory , Nebhan Caroline , Roland Joseph , Weiss Vivian , Johnson Douglas , Richmond Ann 
Published: January 2023

Abstract
Acquired resistance to BRAF/MEK-targeted therapy occurs in the majority of melanoma patients that harbor BRAF-mutated tumors, leading to relapse or progression and the underlying mechanism is unclear in many cases. The tumoral spatial heterogeneity is a potential key to the understanding of the therapy-response relationships, but they also bring challenges of proper parameterization and model validation. In this study, we developed a workflow to detect immune and tumor cells using multiplex immunohistochemistry and spatial imaging analysis from whole-tumor imaging data. Simultaneous characterization of CD11c+, CD8+, CD40+, CD80+ and SOX10+ cells using multispectral imaging was performed on paired tumor sections from eleven melanoma patients prior to BRAF/MEK-targeted therapy and when disease progressed on therapy. We observed a significant increase of tumor cellularity in the progressed tumors, and the close association of SOX10+ melanoma cells with CD8+ cytotoxic T cells (Tc) negatively correlated with patient’s progression-free survival, which may indicate a potential mechanism for acquired resistance to BRAF/MEK-targeted therapy. Using preclinical mouse model of YUMM3.3 BRAF-mutated tumors, we showed the detrimental role of PD-L1:PD-1 axis between melanoma-cell:Tc-cell in melanoma. In TCGA-melanoma dataset (n=445), SOX10hi is associated with a significant reduction of IFNG and GZMB, as well as poor overall survival (OS) in melanoma. Tumor cellularity (SOX10 and MLANA expression) exhibited additive prognostic value in the immune score signature to predict OS in patients with early-stage melanoma and prognoses OS independent of immune score in patients with late-stage melanoma.
This work was supported by a VA Merit Award (I01BX002301) and SRCS award to AR and by R01-CA116021 (AR).
Neutrophil dynamics in the tumor microenvironment determines therapy efficacy and is regulated by microbiota
Authors: Araya Romina E , Lam Khiem C , Huang April , Chen Quanyi , Di Modica Martina , Lopes Amelie , Yang Howard , Liu Huaitian , Lee Maxwell P , Goldszmid Romina S 
Published: January 2023

Abstract
Neutrophils play an important role in cancer progression and both pro- and antitumorigenic functions have been described. However, their role in response to therapy and whether environmental signals modulate their function in the tumor microenvironment (TME) remain unclear. Here, we show that neutrophil content in the TME defines the response to chemo- and immunotherapy. Neutrophil depletion or recruitment blockade impaired the response, while overexpression of neutrophil-attracting chemokines rendered non-responder tumors susceptible to therapy. Importantly, we demonstrate that tumor neutrophils are a heterogeneous and dynamic population that differ from those found in blood or bone marrow, and we identified a neutrophil subset that correlates with therapy response. Additionally, we found that mice lacking microbiota have impaired neutrophil recruitment and function associated with reduced therapy efficacy. Mechanistically, administration of a microbiota-derived NOD2 ligand to mice lacking microbiota was sufficient to restore neutrophil dynamics and response to therapy. We confirmed tumor neutrophil heterogeneity in cancer patients and showed that while total neutrophil content had no predictive value for patient’s overall survival, two specific neutrophil subsets associated with better outcome for pancreatic cancer patients. Remarkably, the subset enriched in non-responder germ-free tumors in our preclinical studies associated with a similar poor outcome in colorectal cancer patients. Collectively, our findings highlight the importance of characterizing neutrophil heterogeneity in the TME and the contribution of microbiota in shaping these cells to predict a successful therapy outcome.
Supported by NIH Intramural Program
Intertumoral CD4+ T-cells instruct monocyte differentiation in pancreatic ductal adenocarcinoma
Authors: Patterson Mike , Burrack Adam L , Schmiechen Zoe C , Xu Yingzheng , Firulyova Maria , Miller Ebony , Schrank Patricia , Zaitsev Konstantin , Williams Jesse , Stromnes Ingunn 
Published: January 2023

Abstract
Pancreatic ductal adenocarcinoma (PDA) is a lethal malignancy that is currently the third leading cause of cancer related deaths. PDA creates a suppressive fibroinflammatory tumor microenvironment (TME) composed of stromal and immune cells that inhibit anti-tumor immune responses. Tumor associated macrophages (TAMs) account for a large percentage of the TME and have remarkably heterogeneous functions, with some subsets promoting anti-tumor responses while others suppress tumor specific T-cells. While many studies have highlighted macrophage heterogeneity in PDA, it remains unclear mechanisms regulating monocyte differentiation into pro vs anti-tumor populations. To understand monocyte differentiation in PDA, we performed trajectory analysis on scRNA-seq data from mouse and human tumors and identified predicted monocyte differentiation pathways toward either MHCII-hi anti-tumor TAMs or Trem2-hi pro-tumor TAMs. Using a newly designed monocyte tracking mouse (CCR2CreER x R26tdTomato) implanted with orthotopic tumors that express the click beetle luciferase (CB) neoantigen, we temporally tracked monocyte differentiation within tumors into TAM populations. Furthermore, using antibody depletion, we found that CD4 depletion led to increased monocyte differentiation into pro-tumor macrophages and a dramatic downregulation of PDL1 on monocyte derived TAMs. Finally, using Ifngr1−/− mice implanted with CB+ tumors, we identify that polarization toward anti-tumor macrophages was driven by IFNy, but TAM PDL1 expression was IFNy independent. Together these data are the first to address monocyte differentiation within PDA and identify previously unexpected role for CD4 T cells governing TAM differentiation program.
The role of the Thrombin/PAR axis in modulating CD8+ T cell anti-tumor immunity
Authors: Cantrell Rachel , Rosenfeldt Leah , Sharma Bal Krishan , Gourley Benjamin , Revenko Alexey , Monia Brett , Palumbo Joseph 
Published: January 2023

Abstract
CD8+ tumor infiltrating lymphocytes (TIL) are critical for tumor clearance following immune checkpoint inhibitor (ICI) therapy. Thrombin, the central hemostatic protease, is activated in the tumor microenvironment and CD8+ T cells express the thrombin-cleavable protease activated receptors (PAR-1, -2, -3, -4). Recent studies showed that thrombin can enhance CD8+ T cell effector functions, but the role of thrombin in ICI is unknown. We hypothesized that thrombin enhances CD8+ TIL anti-tumor effector functions (αTEF) during ICI therapy. Consistent with this hypothesis, we found that reducing circulating prothrombin to ~10% of normal completely abrogated the efficacy of PD-1 blockade therapy to reduce tumor growth, reduced CD8+ TIL infiltration into tumor tissue, and limited αTEF.
Mechanistic in vitro studies showed that thrombin stimulation of isolated murine or human CD8+ T cells resulted in a dose-dependent increase in survival, proliferation and cytokine production (activation profile). Surprisingly, when we stimulated CD8+ T cells lacking PAR-1 or PAR-2, we observed an increase in their activation profile relative to WT CD8+T cells in the absence of thrombin, which was further enhanced with thrombin. Moreover, stimulation of PAR-2 deficient CD8+ T cells with a specific PAR-1 activating peptide resulted in an increased activation profile compared to WT CD8+ T cells stimulated with the same peptide. Together these data suggest that 1) thrombin promotes adaptive tumor immunity in the context of ICI therapy, 2) thrombin promotes αTEF through PAR-1 dependent and independent mechanisms and 3) PAR-2 suppresses CD8+ T cell functions independently of thrombin and limits the capacity of PAR-1 activation to enhance effector functions.
Supported by grants from NIH T32 (5T32AI118697-04)
Utilizing distinct CAR and TCR signaling to generate enhanced cellular immunotherapy
Authors: Gascoigne Nicholas R J , Wu Ling , Brzostek Joanna , Wei Qianru , Vale Previtha Dawn Sakthi , Koh Clara K T , Chua Yen Long , Yap Jiawei , Tan Triscilla Y Y , Lai Junyun , MacAry Paul A 
Published: January 2023

Abstract
CAR-T cells utilize TCR signaling cascades and the recognition functions of antibodies. CAR-T technology has achieved significant success in treatment of certain, primarily liquid, cancers. Nonetheless, many challenges hinder the development of this therapy, for example the efficacy for solid tumors. These challenges show our inadequate understanding of this technology, particularly regarding CAR signaling, and how it may differ from TCR signaling. To dissect CAR signaling, CAR and TCR targeting the same antigen were compared directly. This comparison revealed that CAR was sufficient to bind monomeric antigens due to its high affinity but required oligomeric antigens for its activation. CAR sustained the transduced signal significantly longer than did TCR. CD8 coreceptor was recruited to the CAR synapse but played a negligible role in signaling, unlike for TCR signaling. Surprisingly, we identified a non-canonical CAR signaling triggered in the absence of SRC family kinase (SFK) LCK, which is essential for TCR signaling. We show that LCK-deficient CAR-T cells are strongly signaled through CAR and have a better in vivo efficacy because of reduced exhaustion phenotype and enhanced induction of memory. This non-canonical signaling of CAR-T cells provides new insight into the initiation of TCR and CAR signaling as well as important clinical implications for improvement of CAR function.
Supported by grants from Singapore Ministry of Health’s National Medical Research Council: OFIRG19nov-0066; and Ministry of Education, NUHSRO/2020/110/T1/SEED-MAR/06, and NUS ILO TAP Grant: TAP2002019-04-25. LW, QW and JL were supported by research scholarships from Yong Loo Lin School of Medicine.
Engineering anti-CD229 CAR T cell selectivity for multiple myeloma
Authors: Baker Jillian , Radhakrishnan Sabarinath Venniyil , Olson Michael , Atanackovic Djordje , Luetkens Tim , Mause Erica Vander 
Published: January 2023

Abstract
T cells expressing high affinity chimeric antigen receptors (CAR) against CD229 are able to eradicate multiple myeloma (MM) cells, which express high CD229 levels. However, they also target healthy T cells expressing low CD229 levels. Reducing the affinity of the parental CD229 CAR may increase the selectivity of CD229 CAR T cells, sparing T cells while maintaining activity against MM cells. No robust approaches to engineer minimally altered low affinity variants from existing high-affinity antibodies have been developed.
We generated a site-saturation mutagenesis library of the parental CD229 antibody targeting both CDR3 domains resulting in 305 variants. Variants were expressed as soluble single-chain variable fragments and screened using high-throughput assays to determine binding and expression. Affinities were determined by biolayer interferometry. A subset of 26 binders was further characterized functionally in CAR format using primary human T cells.
Variant CD229 antibodies carrying single amino acid substitutions showed substantially altered expression and binding of recombinant CD229. Affinities of candidate antibodies varied from 150–10,000nM with mainly off-rate driven reductions in affinity. Analyzing the cytotoxic activity of CD229 CAR variants we found that, in contrast to the parental cells, two variant CAR T cells spared normal T cells while maintaining equal cytotoxic activity against MM cells.
Using a novel unbiased mutagenesis approach, we show that generation of minimally altered low affinity variants from existing antibodies is feasible and results in binders with substantially reduced affinity. Low affinity variants show increased selectivity eliminating a key liability of CD229 CAR T cells.
E.R.V received funding from the American Foundation for Pharmaceutical Education (AFPE, Pre-Doctoral Fellowship) and the ALSAM Foundation (Skaggs Fellowship). D.A. received funding from the Huntsman Cancer Institute (HCI, Translational Scholar Award). T.L. received funding from the National Comprehensive Cancer Network (NCCN, Young Investigator Award).
Identifying novel epigenetic modifiers that sensitize T cell malignancies to CD8+ T cell mediated cytotoxicity by high throughput drug screen
Authors: Bradeen Xander Gordon Ray , Christians Allison J , Haverkos Bradley , Davila Eduardo 
Published: January 2023

Abstract
T Cell Lymphomas are a rare subtype of Lymphoma, occurring in ~1% of all cancers and response rates to current therapies reach only ~23%. The lack of unique targetable T cell antigens as well as their tendency to overcome chemotherapies results in a high incidence of relapse in most patients leading to remission rates of approximately two years. Immunotherapies, especially immune-checkpoint blockade, have slightly improved outcomes in refractory disease, but there remains a need for stronger and more durable responses. The chemotherapeutic potential of epigenetic modifiers is just beginning to be explored. HDAC inhibitors, DNA demethylating agents, and histone methyltransferase inhibitors reshape the transcriptome, potentially altering the expression of immune modulatory pathways. We hypothesized that pretreating T cell malignancies with epigenetic modifiers will alter key genetic signatures that sensitize them to CD8+ T cell-mediated cytotoxicity. We pretreated five T cell lymphoma cell lines, representative of different T cell malignant diseases with compounds from an FDA-approved 700+ epigenetic modifying drug library at four different concentrations and then co-cultured them with a tumor-reactive HLA-A2-matched primary (derived from PBMCs) cytotoxic CD8+ T cell line. Changes in T cell mediated cytotoxicity and alterations in immune modulating surface markers and potential CAR-T cell targets (CD30, CCR4, and CD47) were assessed by flow cytometry. Several promising compounds were identified by their ability to improve T cell mediated cytotoxicity, increase expression of immune stimulatory markers, decrease expression of immune inhibitory markers, and augment expression of existing CAR-T target antigens.
Supported by NIH and the Colorado Clinical and Translational Sciences Institute (CCTSI)