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Abstract
Glioblastoma (GBM) is a lethal brain tumor with a 95% five-year mortality rate, and remains a major unmet clinical need. An innovative approach to target GBM is administration of chimeric antigen receptor (CAR) T cells. In this study we identified B7-H3 as a favorable target antigen for CAR T cell therapy of GBM and developed a B7-H3 targeting CAR T cell. This CAR demonstrates robust, antigen-specific cytolysis and cytokine production, low tonic signaling, and preservation of a stem/naïve cell phenotype. Using orthotopic xenograft models of GBM we show that a single intravenous administration of this B7-H3 targeted CAR confers tumor control, and long-term survival. We identified upregulation of T cell inhibitory ligands PD-L1 and PD-L2 as a mechanism of resistance, and modified our B7-H3 targeting CAR vector to include a secreted a PD-1 blocking scFv. We further demonstrate that concomitant PD-1 blockade by the secreted PD-1 blocking scFv augments the antitumor capabilities of B7-H3-targeting CAR T cells through increased intratumoral infiltration by the CAR T cells and improved effector function. These data demonstrate B7-H3 targeting CAR T cells with concomitant PD-1 blockade is a safe and efficient method for improving the antitumor capacity of CAR T cells for GBM.
Authors
Jonathan F. Khan, Parwiz Abrahimi, Winson Cai, Lauren Dong, Kenny Kwok-Hei Yu, Svena Verma, Yacine Marouf, Sadna Budhu, Tara Pal Chaudhuri, Valeria Estrada Navarro, Mamadou A. Bah, Kelsey Hopland, Yanhong Yang, Terence J. Purdon, Ileana C. Miranda, Viviane Tabar, Jedd D. Wolchok, Taha Merghoub, Renier J. Brentjens
Abstract
Authors
Rocio J. Jimenez-Valdes, Quoc Vo, Alison Morris, Chad Steele, Seyed Mehdi Nouraie, Jonathan K. Alder, Kambez H. Benam
Abstract
Although 95% of serotonin in the human body is produced in the gastrointestinal tract, most prior research work has focused on the brain. Despite its abundance in the gut, serotonin’s role in early intestinal development remains poorly understood. Here we show that serotonin is detected in both the small intestine and large intestine as early as 14 to 15 weeks gestational age. Using single-cell transcriptomics with histological validation, we generated a spatiotemporal atlas of serotonin and serotonin-related genes in the human gut from first trimester to late adulthood. The serotonin synthesis gene, TPH1, was exclusively expressed in enteroendocrine cells at all time points analyzed, with the highest expression in the second trimester. The serotonin reuptake gene, SLC6A4, was largely restricted to mature absorptive enterocytes and increased postnatally. Serotonin receptors also displayed distinct cellular and developmental expression patterns. HTR1E was confined to the first trimester during highly proliferative stage. HTR4 was enriched for in crypt regions, suggesting a role in proliferation for serotonin. Finally, HTR3E was associated with tuft cells throughout development. Together, these findings reveal the dynamic, cell-type-specific regulation of serotonin in the developing human gut, suggesting its importance in early intestinal development, and provide a framework to study its roles in intestinal development and diseases.
Authors
Long Phan, Lauren Smith, Kristin M. Milano, Weihong Gu, Madison S. Strine, Harvey J. Kliman, Liza Konnikova
Abstract
Testosterone is a key determinant of red blood cell (RBC) biology and susceptibility to hemolysis; however, its contribution to donor-dependent variation in RBC quality and transfusion effectiveness remains unclear. Using data from 2,357 male participants in the NHLBI REDS-III RBC-Omics study, we quantified plasma total testosterone, free testosterone (bioavailable form), and androstenedione and evaluated their associations with donor characteristics, RBC hemolysis, and recipient post-transfusion hemoglobin increments, a clinical measure of RBC transfusion effectiveness. Subphysiological testosterone concentrations were common (~35%) and independently associated with increased storage and osmotic hemolysis. Frequent blood donations were independently associated with lower circulating free testosterone concentrations, even after adjustment for demographic and hematologic factors. Genome-wide association analyses identified loci associated with circulating testosterone, including regions near SHBG, TNFSF12, and the long noncoding RNA BALR6. Several testosterone variants were associated with RBC transfusion effectiveness. Specifically, variants associated with lower testosterone (e.g., TNFSF12) corresponded to higher post-transfusion hemoglobin increments (+9%), whereas BALR6 variants associated with higher testosterone were linked to reduced hemoglobin increments (–40%). In contrast, androstenedione-associated variants were not related to hemolysis or transfusion outcomes. Together, these findings demonstrate that genetic and physiologic variation in testosterone contributes to inter-individual variability in RBC quality and transfusion effectiveness.
Authors
Fang Fang, Nareg Roubinian, Xutao Deng, Cassie Kemmler, Jost Klawitter, Jelena Klawitter, Touraj Shokati, Uwe Christians, Grier P. Page, Tamir Kanias
Abstract
The role of CD8+ T cells in filarial infections remains poorly understood. Given the chronic nature of these infections, individuals living in endemic regions are frequently exposed to other pathogens including viruses. Because CD8 cells are essential for antiviral immunity, understanding how filarial infections shape the cytotoxic compartment is essential to elucidate their impact on bystander immunity. We evaluated the phenotype and function of CD8+ T cells from Loa loa-infected (Fil+) and -uninfected (Fil–) individuals at baseline and following cytomegalovirus (CMV) re-stimulation. Filarial infection was associated with increased activation and proliferation of CD8+ T cells, characterized by elevated expression of Ki-67, CD107a, and production of type 1 and 17 cytokines. Upon CMV re-stimulation, Fil+ individuals showed markedly reduced expansion of antigen-experienced (CD137+) and polyfunctional (CD137+IFN-γ+TNF-α+, CD137+IFN-γ+TNF-α+IL-2+) CD8+ T cells, along with decreased cytotoxic activity. Unbiased clustering analysis revealed markedly reduced frequency of the CD8+CD45RA+CD57+ subset in Fil+ individuals- a population further defined through transcriptomic profiling that showed enrichment for cytolytic gene signatures (GZMB, GNLY, GZMH, CD244, CX3CR1). These results demonstrated that filarial infection is associated with an altered CD8+ T cell profile that is associated with failure to mount effective viral-specific effector responses, including impaired cytokine production and cytotoxic activity.
Authors
Camila Queiroz-Glauss, Justin Lack, Daniel E. Sturdevant, Thomas B. Nutman
Abstract
The hypothesized cellular and molecular mechanisms underlying dystonia are broad and include mutations that perturb Ca2+ signaling, including those affecting voltage gated calcium channels (VGCC). In mice, pharmacological activation of neuronal L-type VGCCs induces dystonia in a dose dependent manner. Here we demonstrate that mice expressing a gain-of-function mutation in the L-type VGCC CaV1.2, associated with Timothy syndrome (TS), exhibit motor dysfunction consistent with dystonia. Although CaV1.2 is broadly expressed throughout peripheral tissues and across the brain, we establish that the dystonia-like behavior is driven by neuronal expression of the mutant calcium channel and observe an associated potential excitatory/inhibitory (E/I) imbalance. Because patients with TS have profound metabolic dysregulation, which is associated with some dystonias, we measured changes in circulating metabolites. The dystonia-like events are sensitive to perturbations in pyruvate metabolism, reminiscent of a subset of dystonias associated with pyruvate dysregulation. Our study provides insight into the potential convergence of previously established causes of dystonia, calcium signaling and metabolic homeostasis.
Authors
Patrick Towers, Hong-Gang Wang, Maiko Matsui, Geoffrey S. Pitt
Abstract
Pneumocystis jirovecii pneumonia (PCP) remains a major cause of life-threatening respiratory failure in people with advanced HIV, yet the biological factors associated with mortality are incompletely understood. We performed integrated transcriptomic, proteomic, and metabolomic profiling of bronchoalveolar lavage fluid from 42 adults with HIV-associated PCP, relating molecular signatures to mortality. We found that fungal burden, measured by sputum immunofluorescence, Pneumocystis qPCR, and 1,3-β-D-glucan, did not differ between survivors and non-survivors or correlate with baseline gas exchange. Instead, non-survivors exhibited a bronchoalveolar immunometabolic failure signature characterised by marked depletion of immunoglobulins and complement, reduced pattern recognition receptors and fibroblast growth factor family proteins, and broad disruption of extracellular matrix organisation. Metabolomic profiling revealed enrichment of arginine-urea cycle and tricarboxylic acid pathways, with coordinated accumulation of arginine, citrulline, and TCA cycle intermediates consistent with nitrosative and mitochondrial stress. An integrated proteo-metabolomic score summarising this state was significantly higher in non-survivors and did not correlate with fungal burden or cytomegalovirus co-infection. In an immunodeficient precision-cut lung slice model, antigen-screened intravenous immunoglobulin restored macrophage-mediated Pneumocystis clearance, providing proof-of-concept for opsonic augmentation. Our findings indicate that mortality in HIV-PCP is defined by profound immunometabolic failure, highlighting humoral depletion and impaired alveolar repair as potential targets for host-directed adjunctive therapy.
Authors
Peter Rossi-Smith, Ayanda Trevor Mnguni, Dora Pungan, Robert J. Samuels, Vaishnavi R. Kumaran, Leena Syed, Trevor Ferris, Kamil Skirlo, Joseph N. Jarvis, Nelesh P. Govender, Graeme Meintjes, Sean Wasserman, Jay K. Kolls, Rachel P.J. Lai
Abstract
BACKGROUND. Body weight and insulin resistance in women increase in midlife. It is not known whether this is driven by the menopause transition or solely by chronological aging. We aimed to determine the role of the menopause transition and menopause-related BMI changes in the development of insulin resistance in midlife women. METHODS. Mixed effects, linear spline modeling of longitudinal data in 1315 women, 42- to 52-years-old, pre- or early perimenopausal in 1995 to 1996 with up to 16 follow up visits by 2018. Homeostatic-model-assessed-insulin-resistance (HOMA-IR) was measured up to 12 times. Date of final menstrual period (FMP) was prospectively determined. RESULTS. HOMA-IR increased faster over a 4-year interval extending from 1.5 years prior to 2.5 years after the FMP, than in the years before or after it; p value for both slope changes < 0.0001. In the referent woman (White, age 52 years at FMP, non-smoker, body mass index [BMI] 25.6 kg/m2 at baseline, and not on sex hormone therapy or statins), the average annualized rate of HOMA-IR increase, adjusted for covariates including changes in BMI, was 1.2%, 4.3%, and 0.9% respectively, before, during, and after the 4-year transition. CONCLUSION. Consistent with an independent effect of the menopause transition, insulin resistance increased significantly faster during the transition (from 1.5 years before the to 2.5 years after the FMP) than in the years preceding or following the transition. Future studies need to investigate mechanisms underpinning this observed association and determine the impact of lifestyle on dampening menopause-associated increases in insulin resistance. FUNDING. National Institutes of Health.
Authors
Arun S. Karlamangla, Wei Juan Han, Preethi Srikanthan, Albert Shieh, Duncan Thomas, Barbara Sternfeld, Monique M. Hedderson, Gail A. Greendale
Abstract
Chronic liver disease affects over a billion people worldwide. Despite diverse etiologies, a unifying feature of chronic liver disease is the liver’s impaired ability to regenerate during progression to cirrhosis, but no common molecular mechanism has been identified. Hepatocyte proliferation and liver regeneration depend on nucleoplasmic calcium (Ca2+) signals, and Ca2+ signals in hepatocytes depend on the type 2 inositol trisphosphate receptor (ITPR2) calcium release channel. Here, we found that ITPR2 was localized in part to the hepatocyte nucleus, and this localization depended on the presence of nucleoporin 62 (NUP62). Loss of either ITPR2 or NUP62 disrupted nuclear Ca2+ signaling, and impaired Ca2+ signals in the nucleus blunted nuclear entry of β-catenin. Remarkably, both ITPR2 and NUP62 are progressively lost from hepatocytes in patients with the four most common types of chronic liver disease. These findings identify a microdomain regulating Ca2+ signaling in the hepatocyte nucleus that becomes progressively disrupted as liver disease progresses. Preservation of this nuclear microdomain may be a novel approach to maintain liver regeneration and slow the progression to cirrhosis in chronic liver disease.
Authors
Jittima Weerachayaphorn, Mateus T. Guerra, Naotaka Kugiyama, Emma Kruglov, Dejian Zhao, Piyachat Chansela, Vitoon Saengsirisuwan, Marie E. Robert, Teruo Utsumi, Yasuko Iwakiri, Michael H. Nathanson
Abstract
Persistent production of type I interferons (IFN-Is) is a hallmark of cutaneous lupus erythematosus (CLE). Ultraviolet (UV) light stimulates IFN-I response in the skin and exacerbates CLE. Here, we identify V-type immunoglobulin domain-containing suppressor of T cell activation (VISTA) as a negative regulator of both basal and UV-induced IFN-I response in the skin and show that VISTA limits skin photosensitivity in an IFN-I-dependent manner, in part through Stimulator of Interferon Genes (STING). Furthermore, we demonstrate a novel role for VISTA in keratinocytes both at steady state and in response to UV light. Conditional deletion of VISTA in epidermal keratinocytes results in a ~10-fold increase in basal skin IFN-I scores and a heightened UV-induced skin injury score, both of which are dependent on IFN-I signaling. VISTA-targeting monoclonal antibodies suppress the UV-induced IFN-I response in human keratinocytes and in mice expressing human VISTA in vivo, thereby reducing UV-induced skin injury scores. Together, these findings identify VISTA as a keratinocyte-intrinsic checkpoint that restrains STING-associated IFN-I response in the skin and suggest VISTA agonism as a therapeutic strategy to limit photosensitivity in CLE.
Authors
Zachary T. Peters, Lindsay K. Mendyka, J'Voughnn A. Blake, Himanshu B. Goswami, Angelique N. Cortez, Grace E. Crossland, Sicong Shan, Elizabeth C. Nowak, Myana Keusch, Mrinal K. Sarkar, Johann E. Gudjonsson, Christopher M. Burns, Dorothea T. Barton, Bruce R. Blazar, Tyler J. Curiel, Rodwell Mabaera, Victoria P. Werth, Andrea Kalus, Keith B. Elkon, Randolph J. Noelle, Sladjana Skopelja-Gardner
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