bluuebunny/crossref_metadata_2025_split huggingface dataset:
Genetic manipulations to improve intratumoral trafficking of adoptively transferred T cells
Authors: Vianzon Vianca , Garg Ishita , Rogers Laura M 
Published: January 2023

Abstract
T cell–mediated cancer immunotherapies, including anti–PD-1 and CAR-T cells, are becoming standard treatments for many cancer types. One potential strategy to overcome resistance is to enhance the ability of T cells to traffic into tumors. We recently reported results from an innovative, genome-wide screen to identify endogenous T cell genes that enhance intratumoral accumulation using the Sleeping Beauty (SB) transposon system. We successfully identified many promising candidates with this in vivo approach using immunocompetent models of solid tumors.
Others have performed similar independent whole-genome and targeted screens in T cells assessing tissue localization using adoptive T cell transfer approaches. To determine how our gene candidates compared with significant candidates identified in these independent screens, we performed a formal rank aggregation to generate a single, integrated ranked list in descending order of selection strength. Aggregated results yielded approximately 1500 T cell gene candidates whose biological functions are enriched in chemotaxis. We hypothesized that these candidates represent novel therapeutic targets to improve T cell migration into solid tumors.
Here, we present in vivo validation of two of the most strongly selected candidates, including AP2-associated protein kinase 1 (Aak1) and Small Proline-Rich Protein 1B (SPRR1B). Importantly, in vivo migration of adoptively transferred T cells into tumors was significantly increased when the T cells overexpressed AAK1DN125 or SPRR1B. Our observations strongly support the strength of genomic screens to identify truly novel candidates for further development as immunotherapies to enhance T cell homing to the tumor microenvironment.
Supported by Mayo Clinic Graduate School of Biomedical Sciences, K22 (K22CA225786, PI: LM Rogers), and Cancer Immunology and Immunotherapy Program Pilot Award (PI: LM Rogers)
The PARP inhibitor ABT-888 impairs the suppressive capacity of regulatory T cells through modulation of FoxP3 and CTLA-4.
Authors: Falcon Daniel M , Adams Sarah F 
Published: January 2023

Abstract
Immune checkpoint blockade (ICB) has achieved long-term survival in melanoma but results remain limited in other solid malignancies including ovarian cancer. Combination treatments are a promising strategy to optimize responses to ICB. We have previously shown that the PARP inhibitor (PARPi) ABT-888 synergizes with CTLA-4 ICB to achieve long-term survival in a preclinical model of ovarian cancer. While the role of PARPi on DNA repair is well established, the impact on T cells is unclear. Published data using PARP1-KO mice suggests that PARP impairs regulatory T cell (Treg) function through modulation of FoxP3. Tregs suppress conventional T cell responses using multiple mechanisms including consumption of co-stimulatory molecules via the inhibitory receptor CTLA-4. Here we sought to define the impact of ABT-888 on the suppressive function of Tregs using FoxP3-eGFP transgenic mice. Using flow cytometry, we examined the levels of FoxP3 and CTLA-4 in Tregs stimulated in the presence of increasing doses of ABT-888 (0 – 50μM). Independent of stimulation, the GMFI of FoxP3 and CTLA-4 were significantly reduced by ABT-888 in a dose-dependent manner (P &lt; 0.05). To test suppressive function, we pre-treated Tregs with increasing doses of ABT-888 prior to use in a standard proliferation assay. Similarly, ABT-888 pre-treatment significantly reduced the suppressive capacity of Tregs in a dose-dependent manner (P &lt; 0.05). Taken together, these data identify a potentially novel area of synergy between PARPi and ICB. Furthermore, these data highlight the immunomodulatory effects of PARPi which are independent of their role in DNA repair. Future studies will focus on the impact of ABT-888 on tumor-associated Tregs and optimization of ICB therapy.
This research was supported by NCI R37CA229221 (PI: Adams) and UNM Comprehensive Cancer Center Support Grant NCI P30CA11810. D.F. was supported by the Academic Science Education and Research Training program at the University of New Mexico Health Sciences Center (NIGMS Institutional Research and Academic Career Development Award; K12-GM088021).
The novel coreceptor CD8α:MyD88 enhances T-cell proliferation, activation, and antitumor response at low-dose IL-2 in B16 melanoma
Authors: Cao Phoebe , Chen Jiangli , Ciavattone Nicholas , Davila Eduardo 
Published: January 2023

Abstract
The gamma common chain (γc) cytokine receptor plays a central role in Type I cytokine signaling, and in the generation and expansion of antitumor T-cells. Adoptive T-cell therapies (ACT) benefit from IL-2 treatment but is limited by a short half-life and high toxicities. Reducing IL-2 mediated toxicities while retaining antitumor T-cell efficacy is needed to improve clinical outcomes in ACT-treated patients. Other γc cytokines such as IL-7 and 15 have shown benefits for ACT by increasing generation and expansion of long-lived antitumor T-cells with reduced cytokine toxicity. Our lab previously demonstrated that MyD88 (critical adaptor protein in toll-like receptor signaling) is necessary for CD8 T-cell expansion. Induced MyD88 signaling in T-cells via expression of novel coreceptor CD8a:MyD88 (CM T-cells) resulted in enhanced proliferation, activation, and durably reduced tumor size in mouse melanoma models upon ACT treatment. CM T-cells expressed higher γc receptor levels which is required for IL-2, IL-7 and IL-15. Therefore, we hypothesized that CM T-cells have increased sensitivity to γc cytokine signaling resulting in enhanced proliferation, activation, and antitumor activity. CM T cells cultured at 10–25 times lower concentrations of IL-2, -15, -7 demonstrated enhanced cell proliferation and activation and was associated with increased JAK-STAT signaling intensity and persistence. CM T-cell ACT treatment of mice with B16-F10 tumors reduced tumor growth even at the lowest IL-2 concentration (5000 IU IL-2). In conclusion, CM T-cells require lower concentrations of type I cytokines for enhanced antitumor immunity and highlights the potential of CM T-cells to improve clinical outcomes with reduced toxicities.
Supported by grants from NSF GRFP, NCI (R01CA207913, P30CA046934), Dept of VA Merit Award (BX004935-01), University of Colorado Comprehensive Cancer Center (UCCCC), Leukemia and Lymphoma Society Translational Research Program
Tumoral Expression of NAC1 Restrains Antitumor Immunity through the LDHA-mediated Immune Evasion
Authors: Song Jim Jianxun 
Published: January 2023

Abstract
T cell-mediated antitumor immunity has an important role in cancer prevention and treatment; however, immunosuppressive tumor microenvironment (TME) constitutes a major contributor to immune evasion that weakens antitumor immunity. We report here that tumoral expression of nucleus accumbens-associated protein-1 (NAC1), a nuclear factor of the broad complex, tramtrack, bric-a-brac/poxvirus and zinc finger (BTB/POZ) gene family, negatively impacts the CD8+ T cell - mediated antitumor immunity via through LDHA-mediated suppressive tumor microenvironment. We show that NAC1 could positively regulate the expression of lactate dehydrogenase A (LDHA) at transcriptional level, and LDHA-mediated lactic acid accumulation only inhibited cytokine production but also induced the exhaustion and apoptosis of CD8+ cytotoxic T lymphocytes (CTLs), impairing their cell-killing ability. We further show that in the immunocompetent and immunodeficient mice bearing melanoma, tumors with NAC1 depletion grew significantly slower and had elevated infiltration of tumor antigen specific CTLs following adoptive cell transfer, as compared with the control groups. These findings imply that tumor expression of NAC1 contributes substantially to immune evasion through its regulatory role in LDHA expression and lactic production. Thus, therapeutic targeting of NAC1 warrants further exploration as a potential strategy to reinforce cancer immunotherapy such as adoptive cell transfer of CTLs.
Immune characterization in platinum resistant ovarian cancer patients treated with pembrolizumab and guadecitabine
Authors: Xie Ping , Chen Siqi , Cowan Mathew , Huang Hao , Cardenas Horacio , Kocherginsky Masha , Matei Daniela , Zhang Bin 
Published: January 2023

Abstract
Immune checkpoint inhibitors (ICI) have limited activity in ovarian cancer (OC). Accumulating preclinical and early clinical results support that interventions targeting the epigenome could elicit an inflamed tumor milieu to augment ICI. Here we explore the clinical and biological activity of guadecitabine, a second generation Hypomethylating agents (HMA), given in low dose as a priming strategy before pembrolizumab, a humanized anti-PD1 antibody in a clinical trial for patients with recurrent, platinum-resistant OC. Among 35 evaluable patients, there were 3 partial responses (8.6%) and 8 (22.9%) patients with stable disease, resulting in clinical benefit rate (CBR) of 31.4%. Median duration of clinical benefit was 6.8 months. High dimensional immune profiling of PBMCs showed higher frequency of naive and/or central memory CD4+ T cells and of classical monocytes in patients with durable CBR. Higher baseline density of CD8+ T and CD20+ B cells and presence of tertiary lymphoid structures in tumors were associated with durable CBR. The molecular features such as expression levels of PD-L1 or A2AR linked to therapeutic benefit for resistant OC. Our study provides an in-depth view of the immune milieu of platinum resistant OC and of the effects of the combination of HMAs and pembrolizumab on the interactions between immune cell populations and tumor cells.
Supported by USAMRMC/CDMRP (W81XWH170141)
Absence of Toll-like receptor 5 confers survival in mice bearing ovarian tumors treated with anti-PD-L1
Authors: McGinty Mitchell Thomas , Feng Tzu-Yu , Kolli Sree , Putelo Audrey , Rutkowski Melanie 
Published: January 2023

Abstract
Ovarian cancer accounts for more deaths than any other cancer of the female reproductive system. Patients bearing ovarian tumors infiltrated with high frequencies of T cells associate with a greater survival probability. However, therapeutic strategies targeting T cells in ovarian cancers are largely ineffective. We have observed in our models of late-stage murine ovarian cancer that in the absence of Toll-Like Receptor 5 (TLR5) signaling, anti-PD-L1 therapy promotes significant survival and subsequent protection against tumor rechallenge. Ovarian tumors from rechallenged TLR5KO mice exhibited significantly higher frequency/number of both CD4 and CD8 effector memory T cell subsets and greater numbers of antigen experienced T cells. Thus, in the absence of TLR5 signaling, T cells are better able to mount/maintain a response to ovarian tumors after inhibitory receptor blockade. Furthermore, protection against tumor progression in TLR5KO mice corresponds with a significant increase in tumor infiltrating cross presenting and IL-12 producing Dendritic cells (DCs) which could account for the differences in T cell infiltrates. We hypothesize that TLR5 signaling functionally alters DCs to become more suppressive in the tumor microenvironment, impacting T cell differentiation and anti-tumor activity, thereby reducing survival during ovarian tumor progression with aPD-L1 therapy. Clinical implications are already present, as roughly 7.5% of the general population harbor a TLR5 SNP that diminishes TLR5 signaling and is associated with increased long-term survival for ovarian cancer patients. Therefore, patients who express the TLR5 SNP may immediately benefit from anti-PDL1 therapy and those without the SNP, TLR5 antagonism.
Supported by NIH/NCI 1R01CA253285 UVA CIC Collaborative Grant
Nitration of CCL2 disrupts the tumor-protective function of CCL2 in bladder cancer
Authors: Mukherjee Neelam , Ji Niannian , Shu Zhen-Ju , Curiel Tyler J , Svatek Robert S 
Published: January 2023

Abstract
The chemokine CCL2 (C-C motif ligand 2) is best known for its ability to induce trafficking of immune cells by binding its primary receptor, CCR2. The recruitment of immunosuppressive monocytes by CCL2 promotes cancer in several tumor types. Surprisingly, we previously found an unanticipated protective role for CCL2 signaling in bladder cancer (BC). To determine if CCL2’s effect to suppress bladder tumor growth is dependent on T cells, we examined the effects of α-CCL2 antibody in wild-type versus T cell-deficient mice. The protective effect of CCL2 was lost in T cell-deficient mice and on T cell depletion in wild-type mice. Mixed bone marrow chimera experiments also showed that CCR2+ T cells were preferentially recruited by CCL2 to the bladder compared with CCR2− T cells suggesting that the protective effect of CCL2 in BC is through CCR2+ T cells. Most studies researching chemokines, including CCL2, in cancer assume the chemokine to be completely active and do not consider their different modified states which may be one of the critical reasons behind the failure of chemokines in clinical trials. We show for the first time that bladder tumors induce post-translational nitration of CCL2 and block T cell recruitment to the bladder which is restored by exogenous recombinant (r) CCL2 treatment. However, the chemical nitration of rCCL2 abolished this therapeutic efficacy of rCCL2 and was associated with decreased bladder T cell infiltration and more monocytes infiltration in BC. Our study analyzed the role of nitrated CCL2 in BC which explained certain paradoxes between CCL2 levels and patient outcomes in cancer. We also developed a novel BC treatment strategy using rCCL2 which should lead to effective combinations with existing BC immunotherapies.
Supported by (1) the Mays Family Cancer Center at University of Texas Health San Antonio (P30 CA054174), (2) the Roger L. And Laura D. Zeller Charitable Foundation Chair in Urologic Cancer, (3) the Glenda and Gary Woods Distinguished Chair in GU Oncology, (4) the Max & Minnie Tomerlin Voelcker Fund, (5) CDMRP CA170270/P1P2, (6) Bladder Cancer Advocacy Network (BCAN), (7) Research Training Award (RP170345) from the Cancer Prevention & Research Institute of Texas (8) MSTP Program (NIH T32GM113896)
Granzyme F production by CD8 T cells in the tumor microenvironment
Authors: Hay Zachary LeeZer , Knapp Jennifer R , Camp Faye A , O’Connor Brian P , Slansky Jill E 
Published: January 2023

Abstract
Granzymes are a class of cytotoxic proteases and are the primary mechanism utilized by T cells to directly eliminate cancer cells. Each granzyme acts upon a unique set of substrates in target cells to induce cytotoxicity through a range of different mechanisms. Granzymes are some of the most differentially regulated genes in CD8+ tumor infiltrating lymphocytes, relative to T cells outside of the tumor microenvironment (TME). We and others have determined by microarray and qPCR that granzyme F is highly upregulated in the TME. Granzyme F expression is restricted to a small subset of antigen-experienced and exhausted T cells, as determined by flow cytometry-based detection of granzyme F RNA transcripts and may represent a marker of a unique T cell cytotoxic function. Single cell RNA sequencing of CD8 TIL has revealed that granzyme F-high expressing cells are unique from both granzyme A and B expressing cells, and that it is therefore likely these TIL utilize a unique mechanism of cytotoxicity in their elimination of cancer cells. Recombinant granzyme F has previously been shown to induce a unique form of cell death, characterized as being caspase-independent and resulting in rupture of target cell plasma membrane. By over expressing granzyme F we are determining if this mechanism of cell death is leverageable to improve the cytotoxic capacity of TIL, and if induction of different forms of T cell-mediated cytotoxicity can modulate the immunogenicity of the TME. These experiments are designed to provide insight into how to improve adoptive T cell therapies by directly improving cytotoxicity, the terminal step of T cell interaction with tumor cells.
This work was supported by the National Institutes of Health NIAID training grant (Training Program in Immunology; T32-AI07405) award to Zachary Hay
Rebalancing TGFβ1/BMP Signaling Epigenetically Reprograms Fully Exhausted Human CD8 T Cells into a Functional State
Authors: Ghoneim Hazem E , Saadey Abbey A , Yousif Amir , Osborne Nicole , Chen Yu-Lin , Laster Brooke , Zayed Ahmed , Bauman Parker 
Published: January 2023

Abstract
Epigenetic scarring of exhausted T cells (Tex) remains a major obstacle to achieving durable responses by T cell immunotherapies. While we established that de novo DNA methylation programs are causally linked to T cell’s full exhaustion and poor response to immune checkpoint blockade (ICB), major gaps remain in our current understanding of T cell exhaustion—(1) What are the upstream signals that regulate acquisition of exhaustion-specific epigenetic programs? (2) Can we remodel the epigenetic state of Tex to an ICB-responsive state? To address these questions, we developed a novel in vitro model of human T cell dysfunction. First, we demonstrated that chronic TCR stimulation of CD8 T cells is insufficient to develop T cell exhaustion. Instead, our integrative analyses of epigenetic and transcriptional changes in CD8 T cell subsets during chronic virus infections and cancer revealed TGFβ1 as the most significant upstream regulator linked to full exhaustion in mice and humans. Indeed, we found that post-effector TGFβ1 signaling accelerates full exhaustion in chronically stimulated CD8 T cells through stable epigenetic changes linked to impaired effector function and memory potential. Therapeutic rebalancing of TGFβ1/BMP signals by blocking TGFβ1 while boosting BMP signals not only restored effector function, but also unlocked memory programs in human Tex cells. This new therapeutic approach induced a superior anti-tumor activity of human T cells and synergized the ICB response in a murine model of chronic LCMV infection. Our findings highlight the role of TGFβ1/BMP signals in T cell exhaustion and propose a novel therapeutic strategy to epigenetically reprogram fully exhausted T cells, ultimately enhancing T cell immunotherapies.
Sushi domain containing 2 suppresses CD8+ T cell antitumor immunity by targeting IL-2 receptor signaling
Authors: Wen Haitao , Zhao Bao 
Published: January 2023

Abstract
One major mechanism of immunosuppressive tumor microenvironment (TME) is dysfunctional CD8+ T cells which exhibit defective production of antitumor effector molecules. However, the intrinsic molecular mechanism(s) underlying CD8+ T cell dysfunction in cancer remains incompletely understood. Here, we show that the sushi domain containing 2 (SUSD2) is a negative regulator of CD8+ T cell antitumor function. Genetic deletion of Susd2 (Susd2−/−) results in an enhanced production of antitumor molecules in effector CD8+ T cells, which consequently blunts tumor growth in multiple syngeneic mouse tumor models. Through a quantitative mass spectrometry assay, we find that SUSD2 interacts with interleukin-2 receptor a (IL-2Ra) via sushi domain-dependent protein interaction and their interaction suppresses IL-2Ra binding with IL-2, a well-established cytokine essential for CD8+ T cell effector function. Adoptive transfer of Susd2−/− CD19-targeting chimeric antigen receptor (CAR) T cells induces a robust antitumor effect, highlighting SUSD2 as a potential immunotherapy target for cancer treatment.
Supported by grants from NIH (R01 AI62779-01)
Anti-CD123 Chimeric Antigen Receptor Natural Killer cell therapy to treat Acute Myeloid Leukemia.
Authors: Kizerwetter Monika , Yang Huilin , Rahnama Ruyan , Bonifant Challice , Spangler Jamie 
Published: January 2023

Abstract
Acute myeloid leukemia (AML) is a highly prevalent blood and bone marrow cancer characterized by the uncontrolled growth of abnormal myeloblasts. Current treatments for AML often result in systemic toxicities for patients, many of whom will still experience relapse. There is thus an unmet need to develop safer and more effective therapies targeting AML. Chimeric antigen receptor (CAR) T cells, which are engineered to express a cancer-targeting extracellular antibody single-chain variable fragment (scFv) linked to an intracellular T cell-activating domain, have shown promise in treating B cell leukemias, but their success has been limited in myeloid leukemias. This is due in part to the lack of AML-specific target antigens. Moreover, impaired T cell function, a hallmark of AML, is also problematic when relying on autologous T cell activation upon reinfusion. We overcame the limitations of CAR T therapy in AML by developing CAR-expressing natural killer (NK) cells targeting CD123. While present on both healthy and AML cells, CD123 is expressed at &gt;10-fold higher levels on malignant cells. Furthermore, use of NK cells allows use of fully functional allogeneic cells from healthy donors, since NK cells do not induce graft-versus-host-disease. To optimize the tumor cell selectivity of CAR NK cells, we employed the yeast display directed evolution platform to isolate anti-CD123 scFvs with varying affinities and are identifying clones that maximize the interaction with CD123high AML cells over healthy CD123low cells, as determined by binding, cytotoxicity, and cytokine secretion studies. Collectively, our efforts introduce a new paradigm for engineered cell therapeutics with significant promise for AML treatment.
D2C7 CAR: A novel CAR T cell that simultaneously targets wildtype EGFR and its mutant isoform EGFRvIII for treatment of glioma
Authors: Wilkinson Daniel S , Ryan Katherine , Wilson Joseph , Chandramohan Vidyalakshmi , Landi Daniel , Bigner Darell , Fecci Peter E 
Published: January 2023

Abstract
Brain tumors are the leading cause of cancer death in children and a significant cause of morbidity and mortality in adults. Conventional treatments are suboptimal, thus signifying the need for novel therapeutic strategies, such as immunotherapy. Chimeric antigen receptor (CAR) T cells represent a revolutionary class of immunotherapy, achieving considerable success in eliminating hematological cancers but generally failing to control solid tumors in part due to the lack of a suitably-expressed target antigen. Epidermal growth factor receptor (EGFR) is the most ubiquitous and homogeneous antigen on glial brain tumors, and EGFR-directed therapies have been hotly pursued. Moreover, the mutant EGFR variant, EGFRvIII, is present on a subset of pediatric and adult high grade gliomas, representing a targetable, tumor-specific antigen. Unfortunately, CAR T cells targeting EGFRvIII fail to treat tumors possessing as few as 5–10% EGFRvIII-negative cells due to antigen escape. Thus, a CAR T cell that can target both EGFR and EGFRvIII is expected to be superior to a CAR that targets EGFRvIII alone. In this study, we developed a novel third generation CAR T cell consisting of the D2C7scfv targeting moiety that binds a shared epitope between EGFR and EGFRvIII. This D2C7 CAR was able to specifically and potently kill tumor cells expressing wildtype EGFR or EGFRvIII. Importantly, D2C7 CAR significantly prolonged survival of mice bearing EGFR or EGFRvIII-expressing gliomas of both adult (U87) and pediatric (DAOY) origin. Toxicity experiments involving EGFR-expressing human skin grafts provided evidence that D2C7 CAR is safe and effective when administered intracranially to mice bearing intracranial tumors.