Research
Abstract
Type-2 dendritic cells (DC2s) comprise the majority of conventional DCs within most tumors; however, little is known about their ability to initiate and sustain anti-tumor immunity as most studies have focused on antigen cross-presenting Type-1 DCs (DC1s). Here we report that DC2 infiltration identified by analysis of multiple human cancer data sets showed a significant correlation with survival across multiple human cancers, with the benefit being seen in tumors resistant to cytotoxic T cell control. Characterization of DC subtype infiltration into an immunotherapy-resistant model of breast cancer revealed that impairment of DC1s through two unique models resulted in enhanced DC2 functionality and improved tumor control. Batf3-deficiency depleted intratumoral DC1s led to increased DC2 lymph node migration and CD4+ T cell activation. Enhancing DC2 stimulatory potential by genetic deletion of Hsp90b1 (encoding molecular chaperon GP96) led to a similar enhancement of T cell immunity and improved survival in a spontaneous breast cancer model. This data highlights the therapeutic and prognostic potential of DC2s within checkpoint blockade-resistant tumors.
Authors
Stephen Iwanowycz, Soo Ngoi, Yingqi Li, Megan Hill, Christopher Koivisto, Melodie Parrish, Beichu Guo, Zihai Li, Bei Liu
Abstract
Neutrophil-mediated activation and injury of the endothelium play a role in the pathogenesis of diverse disease states ranging from autoimmunity to cancer to COVID-19. Neutralization of cationic proteins (such as neutrophil extracellular trap/NET-derived histones) with polyanionic compounds has been suggested as a potential strategy for protecting the endothelium from such insults. Here, we report that the FDA-approved polyanionic agent defibrotide (a pleotropic mixture of oligonucleotides) directly engages histones and thereby blocks their pathological effects on endothelium. In vitro, defibrotide counteracted endothelial cell activation and pyroptosis-mediated cell death, whether triggered by purified NETs or recombinant histone H4. In vivo, defibrotide stabilized the endothelium and protected against histone-accelerated inferior vena cava thrombosis in mice. Mechanistically, defibrotide demonstrated direct and tight binding to histone H4 as detected by both electrophoretic mobility shift assay and surface plasmon resonance. Taken together, these data provide insights into the potential role of polyanionic compounds in protecting the endothelium from thromboinflammation with potential implications for myriad NET- and histone-accelerated disease states.
Authors
Hui Shi, Alex A. Gandhi, Stephanie A. Smith, Qiuyu Wang, Diane Chiang, Srilakshmi Yalavarthi, Ramadan A. Ali, Chao Liu, Gautam Sule, Pei-Suen Tsou, Yu Zuo, Yogendra Kanthi, Evan A. Farkash, Jiandie D. Lin, James H. Morrissey, Jason S. Knight
Abstract
The alpha ketoglutarate-dependent dioxygenase, prolyl-4-hydroxylase 3 (PHD3), is a Hypoxia-Inducible Factor (HIF) target that uses molecular oxygen to hydroxylate peptidyl prolyl residues. While PHD3 has been reported to influence cancer cell metabolism and liver insulin sensitivity, relatively little is known about effects of this highly conserved enzyme in insulin-secreting β-cells in vivo. Here, we show that deletion of PHD3 specifically in β-cells (βPHD3KO) is associated with impaired glucose homeostasis in mice fed high fat diet. In the early stages of dietary fat excess, βPHD3KO islets energetically rewire, leading to defects in the management of pyruvate fate and a shift from glycolysis to increased fatty acid oxidation (FAO). However, under more prolonged metabolic stress, this switch to preferential FAO in βPHD3KO islets is associated with impaired glucose-stimulated ATP/ADP rises, Ca2+ fluxes and insulin secretion. Thus, PHD3 might be a pivotal component of the β-cell glucose metabolism machinery in mice by suppressing the use of fatty acids as a primary fuel source during the early phases of metabolic stress.
Authors
Daniela Nasteska, Federica Cuozzo, Katrina Viloria, Elspeth M. Johnson, Alpesh Thakker, Rula Bany Bakar, Rebecca L. Westbrook, Jonathan P. Barlow, Monica Hoang, Jamie W. Joseph, Gareth G. Lavery, Ildem Akerman, James Cantley, Leanne Hodson, Daniel A. Tennant, David J. Hodson
Abstract
A dynamically regulated microenvironment, which is mediated by crosstalks between adipocytes 2 and neighboring cells, is critical for adipose tissue homeostasis and function. However, information on 3 key molecules and/or signaling pathways regulating the crosstalks remains limited. In this study, we 4 identify adipocyte miR-182-5p as a crucial anti-obesity molecule that stimulates beige fat thermogenesis 5 by promoting the crosstalk between adipocytes and macrophages. miR-182-5p is highly enriched in 6 thermogenic adipocytes and its expression is markedly stimulated by cold exposure in mice. In contrast, 7 miR-182-5p expression is significantly reduced in adipose tissues of obese humans and mice. Knockout 8 of miR-185-5p decreased cold-induced beige fat thermogenesis whereas overexpression of miR-185-5p 9 increased beiging and thermogenesis in mice. Mechanistically, miR-182-5p promotes FGF21 expression 10 and secretion in adipocytes by suppressing Nr1d1 at 5'UTR, which in turn stimulates acetylcholine 11 synthesis and release in macrophages. Increased acetylcholine expression activates the nicotine 12 acetylcholine receptor in adipocytes, which stimulates PKA signaling and consequent thermogenic gene 13 expression. Our study reveals a key role of the miR-182-5p/FGF21/acetylcholine/acetylcholine receptor 14 axis that mediates the crosstalk between adipocytes and macrophages to promote beige fat 15 thermogenesis. Activation of the miR-182-5p-induced signaling pathway in adipose tissue may be an 16 effective approach to ameliorate obesity and associated metabolic diseases.
Authors
Wen Meng, Ting Xiao, Xiuci Liang, Jie Wen, Xinyi Peng, Jing Wang, Yi Zou, Jiahao Liu, Christie Bialowas, Hairong Luo, Yacheng Zhang, Bilian Liu, Jingjing Zhang, Fang Hu, Meilian Liu, Lily Q. Dong, Zhiguang Zhou, Feng Liu, Juli Bai
Abstract
A major γδ T cell population in human adult blood are the Vγ9Vδ2 T cells that are activated and expanded in a T cell receptor (TCR)-dependent manner by microbe- and endogenous-derived phosphorylated prenyl metabolites (phosphoantigens). Vγ9Vδ2 T cells are also abundant in human fetal peripheral blood, but compared to their adult counterparts they have a distinct developmental origin, are hyporesponsive towards in vitro phosphoantigen exposure and they do not possess a cytotoxic effector phenotype. In order to obtain insight into the role of Vγ9Vδ2 T cells in the human fetus, we investigated their response to in utero infection with the phosphoantigen-producing parasite Toxoplasma gondii (T. gondii). Vγ9Vδ2 T cells expanded strongly in face with congenital T. gondii infection which was associated with differentiation towards potent cytotoxic effector cells. The Vγ9Vδ2 T cell expansion in utero resulted in a fetal footprint with public germline-encoded clonotypes in the Vγ9Vδ2 TCR repertoire 2 months after birth. Overall, our data indicate that the human fetus, from early gestation onwards, possesses public Vγ9Vδ2 T cells that acquire effector functions following parasite infections.
Authors
Ling Ma, Maria Papadopoulou, Martin Taton, Francesca Genco, Arnaud Marchant, Valeria Meroni, David Vermijlen
Abstract
Persistent HPV infection is causative for the majority of cervical cancer (CC) cases; however, current guidelines do not require HPV testing for newly diagnosed CC. Using an institutional cohort of 88 CC patients treated uniformly with standard-of-care chemoradiation (CRT) with prospectively collected clinical outcome data, we observed that patients with cervical tumors containing HPV genotypes other than HPV 16 have worse survival outcomes after CRT compared to patients with HPV 16 positive tumors, consistent with previously published studies. Using RNAseq analysis we quantified viral transcription efficiency and found higher levels of E6 and the alternative transcript E6*I in cervical tumors with HPV genotypes other than HPV 16. These findings were validated using whole transcriptome data from The Cancer Genome Atlas (n=304). For the first time transcript expression level of HPV E6*I was identified as a predictive biomarker of CRT outcome in our complete institutional dataset (n=88), and within the HPV 16 positive subset (n=36). In vitro characterization of HPV E6*I and E6 overexpression revealed that both induce CRT resistance through distinct p53-p21 dependent mechanisms. Our findings suggest that high expression of E6*I and E6 may represent novel biomarkers of CRT efficacy and these patients may benefit from alternative treatment strategies.
Authors
Fiona J. Ruiz, Matthew Inkman, Ramachandran Rashmi, Naoshad Muhammad, Nishanth Gabriel, Christopher A. Miller, Michael D. McLellan, Michael Goldstein, Stephanie Markovina, Perry W. Grigsby, Jin Zhang, Julie K. Schwarz
Abstract
Evidence suggests an association between severe acute respiratory syndrome-cornavirus-2 (SARS-CoV-2) infection and the occurrence of new-onset diabetes. We examined pancreatic expression of angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2), the cell entry factors for SARS-CoV-2, using public single cell RNA sequencing datasets, and pancreas tissue from control male and female non-human primates (NHPs) and humans. We also examined SARS-CoV-2 immunolocalization in pancreas cells of SARS-CoV-2-infected NHPs, and patients deceased from coronavirus disease 2019 (COVID-19). We report expression of ACE2 in pancreatic islet, ductal, and endothelial cells in NHPs and humans. In pancreata from SARS-CoV-2-infected NHPs and COVID-19 patients, SARS-CoV-2 infected ductal, endothelial and islet cells. These pancreata also exhibited generalized fibrosis associated with multiple vascular thrombi. Two out of eight NHPs developed new onset diabetes following SARS-CoV-2 infection. Two out of five COVID-19 patients exhibited new onset diabetes at admission. These results suggest that SARS-CoV-2 infection of the pancreas may promote acute and especially chronic pancreatic dysfunction that could potentially lead to new-onset diabetes.
Authors
Mirza Muhammad Fahd Qadir, Manika Bhondeley, Wandy Beatty, Dina D. Gaupp, Lara A. Doyle-Meyers, Tracy Fischer, Ishitri Bandyopadhyay, Robert V. Blair, Rudolf Bohm, Jay Rappaport, Eric Lazartigues, Richard S. Vander Heide, Jay K. Kolls, Xuebin Qin, Franck Mauvais-Jarvis
Abstract
Some studies suggest that recent common coronavirus (CCV) infections are associated with reduced COVID-19 severity upon SARS-CoV-2 infection. We completed serological assays using samples collected from health care workers to identify antibody types associated with SARS-CoV-2 protection and COVID-19 symptom duration. Rare SARS-CoV-2 cross-reactive antibodies elicited by past CCV infections were not associated with protection; however, the duration of symptoms following SARS-CoV-2 infections was significantly reduced in individuals with higher common betacoronavirus (βCoV) antibody titers. Since antibody titers decline over time after CCV infections, individuals in our cohort with higher βCoV antibody titers were more likely recently infected with common βCoVs compared to individuals with lower antibody titers. Therefore, our data suggest that recent βCoV infections potentially limit the duration of symptoms following SARS-CoV-2 infections through mechanisms that do not involve cross-reactive antibodies. Our data are consistent with the emerging hypothesis that cellular immune responses elicited by recent common βCoV infections transiently reduce symptom duration following SARS-CoV-2 infections.
Authors
Sigrid Gouma, Madison E. Weirick, Marcus J. Bolton, Claudia P. Arevalo, Eileen C. Goodwin, Elizabeth M. Anderson, Christopher M. McAllister, Shannon R. Christensen, Debora Dunbar, Danielle Fiore, Amanda Brock, JoEllen Weaver, John S. Millar, Stephanie DerOhannessian, Ian Frank, Daniel J. Rader, E. John Wherry, Scott E. Hensley
Abstract
Pancreatic ductal adenocarcinoma (PDAC) tumors are characterized by a desmoplastic reaction resulting in dense deposition of collagen that is known to promote cancer progression. A central mediator of pro-tumorigenic collagen signaling is the receptor tyrosine kinase discoid domain receptor 1 (DDR1). DDR1 is a critical driver of a mesenchymal and invasive cancer cell PDAC phenotype. Previous studies have demonstrated that genetic or pharmacologic inhibition of DDR1 reduces PDAC tumorigenesis and metastasis. Here, we investigated whether DDR1 signaling has cancer cell non-autonomous effects that promote PDAC progression and metastasis. We demonstrate that collagen-induced DDR1 activation in cancer cells is a major stimulus for CXCL5 production, resulting in the recruitment of tumor-associated neutrophils (TANs), the formation of neutrophil extracellular traps (NETs) and subsequent cancer cell invasion and metastasis. Moreover, we have identified that collagen-induced CXCL5 production was mediated by a DDR1-PKCθ-SYK-NFkB signaling cascade. Together, these results highlight the critical contribution of collagen I-DDR1 interaction in the formation of an immune microenvironment that promotes PDAC metastasis.
Authors
Jenying Deng, Ya'an Kang, Chien-Chia Cheng, Xinqun Li, Bingbing Dai, Matthew H. Katz, Taoyan Men, Michael P. Kim, Eugene Koay, Huocong Huang, Rolf A. Brekken, Jason B. Fleming
Abstract
Congenital microcephaly (MCPH) is a neurodevelopmental disease associated to mutations in genes encoding proteins involved in centrosomal and chromosomal dynamics during mitosis. Detailed MCPH pathogenesis at the cellular level is still elusive given the diversity of MCPH genes and lack of comparative in vivo studies. By generating a series of CRISPR/Cas9-mediated genetic knockouts we report here that, whereas defects in spindle pole proteins (ASPM, MCPH5) result in mild microcephaly during development, lack of centrosome (CDK5RAP2, MCPH3) or centriole (CEP135, MCPH8) regulators induces delayed chromosome segregation and chromosomal instability in neural progenitors (NPs). Our novel mouse model of MCPH8 suggests that Cep135 deficiency results in centriole duplication, TP53 activation and cell death of NPs. Trp53 ablation in a Cep135-deficient background prevents cell death, but not microcephaly, and leads to subcortical heterotopias, a malformation seen in MCPH8 patients. These results suggest that microcephaly in some MCPH patients can arise from the lack of adaptation to centriole defects in NPs and may lead to architectural defects if chromosomally unstable cells are not eliminated during brain development.
Authors
José González-Martínez, Andrzej W. Cwetsch, Diego Martínez-Alonso, Luis R. López-Sainz, Jorge Almagro, Anna Melati, Jesús Gómez, Manuel Pérez-Martínez, Diego Megías, Jasminka Boskovic, Javier Gilabert-Juan, Osvaldo Graña Castro, Alessandra Pierani, Axel Behrens, Sagrario Ortega, Marcos Malumbres
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