Children and adults with Philadelphia chromosome-like B cell acute lymphoblastic leukemia (Ph-like B-ALL) experience high relapse rates despite best-available conventional chemotherapy. Ph-like ALL is driven by genetic alterations that activate constitutive cytokine receptor and kinase signaling, and early-phase trials are investigating the potential of tyrosine kinase inhibitor (TKI) addition to chemotherapy to improve clinical outcomes. However, preclinical studies have shown that JAK or PI3K pathway inhibition is insufficient to eradicate the most common cytokine receptor-like factor 2 (CRLF2)-rearranged Ph-like ALL subset. We thus sought to define additional essential signaling pathways required in Ph-like leukemogenesis for improved therapeutic targeting. Herein, we describe a novel adaptive signaling plasticity of CRLF2-rearranged Ph-like ALL following selective TKI pressure, which occurs in the absence of genetic mutations. Interestingly, we observed that Ph-like ALL cells have activated SRC, ERK and PI3K signaling consistent with activated B-cell receptor (BCR) signaling, although they do not express cell surface mu heavy chain (uHC). Combinatorial targeting of JAK/STAT, PI3K, and ‘BCR-like’ signaling with multiple TKIs and/or dexamethasone prevented this signaling plasticity and induced complete cell death, demonstrating a more optimal and clinically pragmatic therapeutic strategy for CRLF2-rearranged Ph-like ALL.
Christian Hurtz, Gerald B. Wertheim, Joseph P. Loftus, Daniel Blumenthal, Anne Lehman, Yong Li, Asen Bagashev, Bryan Manning, Katherine D. Cummins, Janis K. Burkhardt, Alexander E. Perl, Martin Carroll, Sarah K. Tasian
T follicular helper (Tfh) cells are indispensable for the formation of germinal center (GC) reactions, while T follicular regulatory (Tfr) cells inhibit Tfh-mediated GC responses. Aberrant activation of Tfh cells contributes significantly to the pathogenesis of autoimmune diseases, such as systemic lupus erythematosus (SLE). Nonetheless, the molecular mechanisms mitigating excessive Tfh cell differentiation, which in turn trigger autoimmunity, are not fully understood. Herein we demonstrate that the adenovirus E4 promoter-binding protein (E4BP4) mediates a feedback loop and acts as a transcriptional brake to inhibit Tfh cell differentiation. Furthermore, we show that such an immunological mechanism is compromised in patients with SLE. Establishing mice with either conditional knock-out (cKO) or knock-in (cKI) of the E4bp4 gene in T cells reveals that E4BP4 strongly inhibits Tfh cell differentiation. Mechanistically, E4BP4 deregulates Bcl6 transcription by recruiting the repressive epigenetic modifiers HDAC1 and EZH2. E4BP4 phosphorylation site mutants had limited capability with regard to inhibiting Tfh cell differentiation. In SLE, we detected impaired phosphorylation of E4BP4, finding that this compromised transcription factor is positively correlated with disease activity. These findings unveiled molecular mechanisms by which E4BP4 restrains Tfh cell differentiation, whose compromised function is associated with uncontrolled autoimmune reactions in SLE.
Zijun Wang, Ming Zhao, Jinghua Yin, Limin Liu, Longyuan Hu, Yi Huang, Aiyun Liu, Jiajun Ouyang, Xiaoli Min, Shijia Rao, Wenhui Zhou, Haijing Wu, Akihiko Yoshimura, Qianjin Lu
Proliferation of CD4+ T cells harboring HIV-1 proviruses is a major contributor to viral persistence in people on antiretroviral therapy (ART). To determine whether differential rates of clonal proliferation or HIV-1-specific CTL pressure shape the provirus landscape, we performed the intact proviral DNA assay (IPDA) and obtained 661 near-full length provirus sequences from eight individuals with suppressed viral loads on ART at time points seven years apart. We observed slow decay of intact proviruses but no changes in the proportions of various types of defective proviruses. The proportion of intact proviruses in expanded clones was similar to that of defective proviruses in clones. Intact proviruses observed in clones did not have more escaped CTL epitopes than intact proviruses observed as singlets. Concordantly, total proviruses at later timepoints or observed in clones were not enriched in escaped or unrecognized epitopes. Three individuals with natural control of HIV-1 infection (controllers) on ART, included because controllers have strong HIV-1-specific CTL responses, had a smaller proportion of intact proviruses but a similar distribution of defective provirus types and escaped or unrecognized epitopes as the other individuals. This work suggests that CTL selection does not significantly check clonal proliferation of infected cells or greatly alter the provirus landscape in people on ART.
Annukka A. R. Antar, Katharine M. Jenike, Sunyoung Jang, Danielle N. Rigau, Daniel B. Reeves, Rebecca Hoh, Melissa R. Krone, Jeanne C. Keruly, Richard D. Moore, Joshua T. Schiffer, Bareng A.S. Nonyane, Frederick M. Hecht, Steven G. Deeks, Janet D. Siliciano, Ya-Chi Ho, Robert F. Siliciano
Background. Post-receptor insulin resistance (IR) is associated with hyperglycemia and hepatic steatosis. However, receptor-level IR (e.g. insulin receptor pathogenic variants, INSR) causes hyperglycemia without steatosis. We examined four pathologic conditions of IR in humans to examine pathways controlling lipid metabolism and gluconeogenesis. Methods. Cross-sectional study of severe, receptor IR (INSR, n=7), versus post-receptor IR that was severe (lipodystrophy, n=14), moderate (type 2 diabetes [T2D], n=9) or mild (obesity, n=8). Lipolysis (glycerol turnover), hepatic glucose production (HGP), gluconeogenesis (deuterium incorporation from body water into glucose), hepatic triglyceride (magnetic resonance spectroscopy), and hepatic fat oxidation (plasma β-hydroxybutyrate) were measured. Results. Lipolysis was 2-3-fold higher in INSR versus all other groups, and HGP 2-fold higher in INSR and lipodystrophy versus T2D and obesity (p<0.001) suggesting severe adipose and hepatic IR. INSR subjects had a higher contribution of gluconeogenesis to HGP, ~77%, versus 52-59% in other groups (p=0.0001). Despite high lipolysis, INSR subjects had low hepatic triglycerides (0.5 [0.1-0.5]), in contrast to lipodystrophy (10.6 [2.8-17.1], p<0.0001). β-hydroxybutyrate was 2-7-fold higher in INSR versus all other groups (p<0.0001) consistent with higher hepatic fat oxidation. Conclusion. These data support a key pathogenic role of adipose tissue IR to increase glycerol and FFA availability to the liver in both receptor and post-receptor IR. However, the fate of FFA diverges in these populations. In receptor-level IR, FFA oxidation drives gluconeogenesis rather than being reesterified to triglyceride. In contrast, in post-receptor IR, FFA contributes to both gluconeogenesis and hepatic steatosis. Trial registration. ClinicalTrials.gov NCT01778556; NCT00001987; NCT02457897 Funding. NIDDK, USDA ARS 58-3092-5-001
Hilal Sekizkardes, Stephanie T. Chung, Shaji Chacko, Morey Haymond, Megan Startzell, Mary Walter, Peter J. Walter, Marissa Lightbourne, Rebecca J. Brown
The atypical cadherin FAT4 has established roles in regulation of planar cell polarity and Hippo pathway signaling that are cell context dependent. The recent identification of FAT4 mutations in Hennekam syndrome, features of which include lymphedema, lymphangiectasia and mental retardation, uncovered an important role for FAT4 in the lymphatic vasculature. Hennekam syndrome is also caused by mutations in CCBE1 and ADAMTS3, encoding a matrix protein and protease, respectively, that regulate activity of the key pro-lymphangiogenic VEGF-C/VEGFR3 signaling axis by facilitating the proteolytic cleavage and activation of VEGF-C. The fact that FAT4, CCBE1 and ADAMTS3 mutations underlie Hennekam syndrome suggested that all three genes might function in a common pathway. We identified FAT4 as a target gene of GATA2, a key transcriptional regulator of lymphatic vascular development and in particular, lymphatic vessel valve development. Here, we demonstrate that FAT4 functions in a lymphatic endothelial cell autonomous manner to control cell polarity in response to flow and is required for lymphatic vessel morphogenesis throughout development. Our data reveal a crucial role for FAT4 in lymphangiogenesis and shed light on the mechanistic basis by which FAT4 mutations underlie a human lymphedema syndrome.
Kelly L. Betterman, Drew L. Sutton, Genevieve A. Secker, Jan Kazenwadel, Anna Oszmiana, Lillian Lim, Naoyuki Miura, Lydia Sorokin, Benjamin M. Hogan, Mark L. Kahn, Helen McNeill, Natasha L. Harvey
Chronic inflammation is a pathologic feature of neurodegeneration and aging; however, the mechanism regulating this process is not understood. Melatonin, an endogenous free radical scavenger synthesized by neuronal mitochondria, decreases with aging and neurodegeneration. We proposed that insufficient melatonin levels impair mitochondrial homeostasis resulting in mitochondrial DNA (mtDNA) release, activation of cytosolic DNA mediated inflammatory response in neurons. We found increased mitochondrial oxidative stress and decreased mitochondrial membrane potential with higher mitochondrial DNA (mtDNA) release in brain and primary cerebro-cortical neurons of melatonin deficient aralkylamine N-acetyltransferase (AANAT) knockout mice. Cytosolic mtDNA activated the cGAS/STING/IRF3 pathway, stimulating inflammatory cytokine generation. We found that Huntington's disease mice increased mtDNA release, cGAS activation, and inflammation, all inhibited by exogenous melatonin. Thus, we demonstrated that cytosolic mtDNA activated the inflammatory response in aging and neurodegeneration, a process modulated by melatonin. Furthermore, our data suggest that AANAT knockout mice are a model of accelerated aging.
Abhishek Jauhari, Sergei V. Baranov, Yalikun Suofu, Jinho Kim, Tanisha Singh, Svitlana Yablonska, Fang Li, Xiaomin Wang, Patrick Oberly, M. Beth Minnigh, Samuel M. Poloyac, Diane L. Carlisle, Robert M. Friedlander
Multiple sclerosis (MS) is an inflammatory demyelinating disorder of the CNS. Bile acids are cholesterol metabolites that can signal through receptors on cells throughout the body, including the CNS and immune system. Whether bile acid metabolism is abnormal in MS is unknown. Using global and targeted metabolomic profiling, we identified lower levels of circulating bile acid metabolites in multiple cohorts of adult and pediatric MS patients compared to controls. In white matter lesions from MS brain tissue, we noted the presence of bile acid receptors on immune and glial cells. To mechanistically examine the implications of lower levels of bile acids in MS, we studied the in vitro effects of an endogenous bile acid – tauroursodeoxycholic acid (TUDCA) on astrocyte and microglial polarization. TUDCA prevented neurotoxic (A1) polarization of astrocytes and pro-inflammatory polarization of microglia in a dose-dependent manner. TUDCA supplementation in experimental autoimmune encephalomyelitis reduced severity of disease through its effects on GPBAR1, based on behavioral and pathological measures. We demonstrate that bile acid metabolism is altered in MS; bile acid supplementation prevents polarization of astrocytes and microglia to neurotoxic phenotypes and ameliorates neuropathology in an animal model of MS. These findings identify dysregulated bile acid metabolism as a potential therapeutic target in MS.
Pavan Bhargava, Matthew D. Smith, Leah Mische, Emily P. Harrington, Kathryn C. Fitzgerald, Kyle A. Martin, Sol Kim, Arthur Anthony A. Reyes, Jaime Gonzalez-Cardona, Christina Volsko, Ajai Tripathi, Sonal Singh, Kesava Varanasi, Hannah-Noelle Lord, Keya R. Meyers, Michelle Taylor, Marjan Gharagozloo, Elias S. Sotirchos, Bardia Nourbakhsh, Ranjan Dutta, Ellen Mowry, Emmanuelle Waubant, Peter A. Calabresi
β-cell apoptosis and dedifferentiation are two hotly-debated mechanisms underlying β-cell loss in type 2 diabetes; however, the molecular drivers underlying such events remain largely unclear. Here, we performed a side-by-side comparison of mice carrying β-cell-specific deletion of endoplasmic reticulum (ER)-associated degradation (ERAD) and autophagy. We reported that while autophagy was necessary for β-cell survival, the highly conserved Sel1L-Hrd1 ERAD protein complex was required for the maintenance of β-cell maturation and identity. Using single cell RNA-sequencing, we demonstrated that Sel1L deficiency was not associated with β-cell loss, but rather loss of β-cell identity. Sel1L-Hrd1 ERAD controlled β-cell identity via TGFβ signaling, in part by mediating the degradation of TGFβ receptor 1 (TGFβRI). Inhibition of TGFβ signaling in Sel1L-deficient β-cells augmented the expression of β-cell maturation markers and increased the total insulin content. Our data revealed distinct pathogenic effects of two major proteolytic pathways in β-cells, providing a new framework for therapies targeting distinct mechanisms of protein quality control.
Neha Shrestha, Tongyu Liu, Yewei Ji, Rachel Reinert, Mauricio Torres, Xin Li, Maria Zhang, Chih-Hang Anthony Tang, Chih-Chi Andrew Hu, Chengyang Liu, Ali Naji, Ming Liu, Jiandie D. Lin, Sander Kersten, Peter Arvan, Ling Qi
Plasmacytoid dendritic cells (pDC), the major producers of Type I interferon, are principally recognized as key mediators of antiviral immunity. However, their role in tumor immunity is less clear. Depending on the context, pDC can both promote or suppress antitumor immune responses. In this study, we identified a naturally occurring pDC subset expressing high levels of OX40 (OX40+ pDC) enriched in the tumor microenvironment (TME) of head and neck squamous cell carcinoma. OX40+ pDC were distinguished by a distinct immunostimulatory phenotype, cytolytic function and ability to synergize with conventional dendritic cells (cDC) in generating potent tumor antigen-specific CD8+ T cell responses. Transcriptomically, we found they selectively utilized EIF2 signaling and oxidative phosphorylation pathways. Moreover, depletion of pDC in the murine OX40+ pDC-rich tumor model accelerated tumor growth. Collectively, we present evidence of a pDC subset in the TME that favors antitumor immunity.
Kate O. Poropatich, Donye Dominguez, Wen-Ching Chan, Jorge Andrade, Yuanyuan Zha, Brian D. Wray, Jason Miska, Lei Qin, Lisa E. Cole, Sydney Coates, Urjeet A. Patel, Sandeep Samant, Bin Zhang
Glioblastoma (GBM) contains a subpopulation of cells, GBM stem cells (GSCs), that maintain the bulk tumor and represent a key therapeutic target. Norrin is a Wnt ligand that binds the Frizzled4 (FZD4) receptor to activate canonical Wnt signaling. While Norrin, encoded by NDP, has a well- described role in vascular development, its function in human tumorigenesis is largely unexplored. Here, we show that NDP expression is enriched in neurological cancers, including GBM, and its levels positively correlated with survival in a GBM subtype defined by low expression of ASCL1, a proneural factor. We investigated the function of Norrin and FZD4 in GSCs and found that it mediated opposing tumor-promoting and -suppressive effects on ASCL1lo and ASCL1hi GSCs. Consistent with a potential tumor suppressive effect of Norrin suggested by the tumour outcome data, we found that Norrin signaling through FZD4 inhibited growth in ASCL1lo GSCs. In contrast, in ASCL1hi GSCs Norrin promoted Notch signaling, independently of WNT, to promote tumor progression. Forced ASCL1 expression reversed the tumor suppressive effects of Norrin in ASCL1lo GSCs. Our results identify Norrin as a modulator of human brain cancer progression and reveal an unanticipated Notch mediated function of Norrin in regulating cancer stem cell biology.
Ahmed El-Sehemy, Hayden J. Selvadurai, Arturo Ortin-Martinez, Neno T. Pokrajac, Yasin Mamatjan, Nobuhiko Tachibana, Katherine J. Rowland, Lilian Lee, Nicole I. Park, Kenneth D. Aldape, Peter Dirks, Valerie A. Wallace
Infusion of the broadly neutralizing antibody VRC01 has been evaluated in HIV-1 chronically infected individuals. Here we studied how VRC01 infusions impacted viral rebound after cessation of antiretroviral therapy (ART) in 18 acutely-treated and durably-suppressed individuals. Viral rebound occurred in all individuals, yet VRC01 infusions modestly delayed rebound and participants who showed a faster decay of VRC01 in serum rebounded more rapidly (Rho=0.60, p=0.03). Participants with strains most sensitive to VRC01 or with VRC01 epitope motifs similar to known VRC01-susceptible strains rebounded later (Rho=-0.70, p<0.03). Upon rebound, HIV-1 sequences were indistinguishable from those sampled at diagnosis. Across the cohort, participant derived Env showed different sensitivity to VRC01 neutralization (including two resistant viruses), yet neutralization sensitivity was similar at diagnosis and post-rebound, indicating the lack of selection for VRC01-resistance during treatment interruption.Our results showed that viremia rebounded despite the absence of HIV-1 adaptation to VRC01 and an average VRC01 trough of 221µg/mL. While VRC01 levels were insufficient to prevent a resurgent infection, knowledge that they did not mediate Env mutations in acute-like viruses is relevant for antibody-based strategies in acute infection.
Evan M. Cale, Hongjun Bai, Meera Bose, Michael A. Messina, Donn Colby, Eric Sanders-Buell, Bethany L. Dearlove, Yifan Li, Emily Engeman, Daniel Silas, Anne Marie O’Sullivan, Brendan Mann, Suteeraporn Pinyakorn, Jintana Intasan, Khunthalee Benjapornpong, Carlo Sacdalan, Eugene Kroon, Nittaya Phanuphak, Robert Gramzinski, Sandhya Vasan, Merlin L. Robb, Nelson L. Michael, Rebecca M. Lynch, Robert Bailer, Amélie Pagliuzza, Nicolas Chomont, Amarendra Pegu, Nicole A. Doria-Rose, Lydie Trautmann, Trevor A. Crowell, John Mascola, Jintanat Ananworanich, Sodsai Tovanabutra, Morgane Rolland
In patients with HBV and HCV coinfection, HBV reactivation leading to severe hepatitis has been reported with the use of direct-acting antivirals (DAAs) to treat HCV infection. Here we study the molecular mechanisms behind this viral interaction. In coinfected cell culture and humanized mice, HBV replication was suppressed by HCV coinfection. In vitro, HBV suppression was attenuated when interferon signaling was blocked. In vivo, HBV viremia, after initial suppression by HCV super-infection, rebounded following HCV clearance by DAA treatment that was accompanied by a reduced hepatic interferon response. Using blood samples of coinfected patients, interferon-stimulated gene products including C-X-C motif chemokine 10 (CXCL10) and C-C motif chemokine ligand 5 (CCL5), and alanine aminotransferase (ALT) were identified to have predictive value for HBV reactivation after HCV clearance. Taken together, our data suggest that HBV reactivation is a result of diminished hepatic interferon response following HCV clearance and identifies serologic markers that can predict HBV reactivation in DAA-treated HBV-HCV coinfected persons.
Xiaoming Cheng, Takuro Uchida, Yuchen Xia, Regina Umarova, Chun-Jen Liu, Pei-Jer Chen, Anuj Gaggar, Vithika Suri, Marcus Maximilian Mücke, Johannes Vermehren, Stefan Zeuzem, Yuji Teraoka, Mitsutaka Osawa, Hiroshi Aikata, Keiji Tsuji, Nami Mori, Shuhei Hige, Yoshiyasu Karino, Michio Imamura, Kazuaki Chayama, T. Jake Liang
As there is growing evidence for the tumor microenvironment’s (TME) role in tumorigenesis, we investigated the role of fibroblast-expressed kinases in triple negative breast cancer (TNBC). Using a high-throughput kinome screen combined with 3D invasion assays, we identified fibroblast-expressed PIK3Cδ (f-PIK3Cδ) as a key regulator of progression. Although PIK3Cδ was expressed in primary fibroblasts derived from TNBC patients, it was undetectable in breast cancer cell lines. Genetic and pharmacologic gain- and loss-of functions experiments verified the contribution of f-PIK3Cδ in TNBC cell invasion. Integrated secretomics and transcriptomics analyses revealed a paracrine mechanism via which f-PIK3Cδ confers its pro-tumorigenic effects. Inhibition of f-PIK3Cδ promoted the secretion of factors, including PLGF and BDNF, which led to upregulation of NR4A1 in TNBC cells where it acts as a tumor suppressor. Inhibition of PIK3Cδ in an orthotopic BC mouse model reduced tumor growth only after inoculation with fibroblasts, indicating a role of f-PIK3Cδ in cancer progression. Similar results were observed in the MMTV-PyMT transgenic BC mouse model, along with a decrease on tumor metastasis emphasizing the potential immune-independent effects of PIK3Cδ inhibition. Finally, analysis of BC patient cohorts and TCGA datasets identified f-PIK3Cδ (protein and mRNA levels) as an independent prognostic factor for overall and disease free survival, highlighting it as a therapeutic target for TNBC.
Teresa Gagliano, Kalpit Shah, Sofia Gargani, Liyan Lao, Mansour Alsaleem, Jianing Chen, Vasileios Ntafis, Penghan Huang, Angeliki Ditsiou, Viviana Vella, Kritika Yadav, Kamila Bienkowska, Giulia Bresciani, Kai Kang, Leping Li, Philip Carter, Graeme Benstead-Hume, Timothy O’Hanlon, Michael Dean, Frances M.G. Pearl, Soo Chin Lee, Emad A. Rakha, Andrew R Green, Dimitris L. Kontoyiannis, Erwei Song, Justin Stebbing, Georgios Giamas
As treatment of the early, inflammatory phase of sepsis improves, post-sepsis immunosuppression and secondary infection have increased in importance. How early inflammation drives immunosuppression remains unclear. Although IFNγ typically helps microbial clearance, we found that increased plasma IFNγ in early clinical sepsis was associated with the later development of secondary Candida infection. Consistent with this observation, we found that exogenous IFNγ suppressed macrophage phagocytosis of zymosan in vivo, and antibody blockade of IFNγ after endotoxemia improved survival of secondary candidemia. Transcriptomic analysis of innate lymphocytes during endotoxemia suggested that NKT cells drove IFNγ production by NK cells via mTORC1. Activation of iNKT cells with glycolipid antigen drove immunosuppression. Deletion of iNKT cells in Cd1d-/- mice or inhibition of mTOR by rapamycin reduced immunosuppression and susceptibility to secondary Candida infection. Thus, although rapamycin is typically an immunosuppressive medication, in the context of sepsis, rapamycin has the opposite effect. These results implicated a NKT cell-mTOR-IFNγ axis in immunosuppression following endotoxemia or sepsis. In summary, in vivo iNKT cells activated mTORC1 in NK cells to produce IFNγ , which worsened macrophage phagocytosis, clearance of secondary Candida infection and mortality.
Edy Y. Kim, Hadas Ner-Gaon, Jack Varon, Aidan M. Cullen, Jingyu Guo, Jiyoung Choi, Diana Barragan-Bradford, Angelica Higuera, Mayra Pinilla-Vera, Samuel A.P. Short, Antonio J. Arciniegas-Rubio, Tomoyoshi Tamura, David E. Leaf, Rebecca M. Baron, Tal Shay, Michael B. Brenner
Kallikrein-related peptidase 6 (KLK6) is a secreted serine protease hypothesized to promote inflammation via cleavage of protease-activated receptors (PAR)1 and PAR2. KLK6 levels are elevated in multiple inflammatory and autoimmune conditions, but no definitive role in pathogenesis has been established. Here, we show that skin-targeted overexpression of KLK6 causes generalized, severe psoriasiform dermatitis with spontaneous development of debilitating psoriatic arthritis-like joint disease. The psoriatic skin and joint phenotypes are reversed by normalization of skin KLK6 levels and attenuated following genetic elimination of PAR1 but not PAR2. Conservation of this regulatory pathway was confirmed in human psoriasis using vorapaxar, an FDA-approved PAR1 antagonist, on explanted lesional skin from psoriasis patients. Beyond defining a critical role for KLK6-PAR1 signaling in promoting psoriasis, our results demonstrate that KLK6-PAR1-mediated inflammation in the skin alone is sufficient to drive inflammatory joint disease. Further, we identify PAR1 as a promising cytokine-independent target in therapy of psoriasis and psoriatic arthritis.
Allison C. Billi, Jessica E. Ludwig, Yi Fritz, Richard Rozic, William R. Swindell, Lam C. Tsoi, Dennis Gruszka, Shahla Abdollahi-Roodsaz, Xianying Xing, Doina Diaconu, Ranjitha Uppala, Maya I. Camhi, Philip A. Klenotic, Mrinal K. Sarkar, M. Elaine Husni, Jose U. Scher, Christine McDonald, J. Michelle Kahlenberg, Ronald J. Midura, Johann E. Gudjonsson, Nicole L. Ward
To identify neurons that specifically increase blood glucose from among the diversely-functioning cell types in the ventromedial hypothalamic nucleus (VMN), we studied the cholecystokinin (CCK) receptor-B (CCKBR)-expressing VMN targets of glucose-elevating parabrachial nucleus neurons. Activating these VMNCCKBR neurons increased blood glucose. Furthermore, while silencing the broader VMN decreased energy expenditure and promoted weight gain without altering blood glucose, silencing VMNCCKBR neurons decreased hepatic glucose production (HGP), insulin-independently decreasing blood glucose without altering energy balance. Silencing VMNCCKBR neurons also impaired the counter-regulatory response (CRR) to insulin-induced hypoglycemia and glucoprivation and replicated hypoglycemia-associated autonomic failure (HAAF). Hence, VMNCCKBR cells represent a specialized subset of VMN cells that function to elevate glucose. These cells not only mediate the allostatic response to hypoglycemia, but also insulin-independently modulate the homeostatic setpoint for blood glucose, consistent with a role for the brain in the insulin-independent control of glucose homeostasis.
Jonathan N. Flak, Paulette Goforth, James Dell'Orco, Paul V. Sabatini, Chien Li, Nadejda Bozadjieva, Matthew J. Sorensen, Alec C. Valenta, Alan C. Rupp, Alison H. Affinati, Corentin Cras-Méneur, Ahsan Ansari, Jamie Sacksner, Nandan Kodur, Darleen A. Sandoval, Robert t. Kennedy, David Olson, Martin G. Myers Jr.
Neutrophil accumulation is associated with lung pathology during active tuberculosis (ATB). However, the molecular mechanism(s) by which neutrophils accumulate in the lung and contribute to TB immunopathology is not fully delineated. Using the well-established mouse model of TB, our new data provides evidence that the alarmin S100A8/A9 mediates neutrophil accumulation during progression to chronic TB. Depletion of neutrophils or S100A8/A9 deficiency resulted in improved Mycobacterium tuberculosis (Mtb) control during chronic but not acute TB. Mechanistically, we demonstrate that following Mtb infection, S100A8/A9 expression is required for upregulation of the integrin molecule CD11b specifically on neutrophils, mediating their accumulation during chronic TB disease. These findings are further substantiated by increased expression of S100A8 and S100A9 mRNA in whole blood in human TB progressors when compared to non-progressors, and rapidly decreased S100A8/A9 protein levels in the serum upon TB treatment. Furthermore, we demonstrate that S100A8/A9 serum levels along with chemokines are useful in distinguishing between ATB and asymptomatic Mtb-infected latent individuals. Thus, our results support targeting S100A8/A9 pathways as host-directed therapy for TB.
Ninecia R. Scott, Rosemary V. Swanson, Noor Al-Hammadi, Racquel Domingo-Gonzalez, Javier Rangel-Moreno, Belinda A. Kriel, Allison N. Bucsan, Shibali Das, Mushtaq Ahmed, Smriti Mehra, Puthayalai Treerat, Alfredo Cruz-Lagunas, Luis Jimenez-Alvarez, Marcela Muñoz-Torrico, Karen Bobadilla-Lozoya, Thomas Vogl, Gerhard Walzl, Nelita du Plessis, Deepak Kaushal, Thomas Scriba, Joaquin Zuñiga, Shabaana Khader
PD-1 expression is a hallmark of both early antigen-specific T-cell activation and later chronic stimulation suggesting key roles in both naive T-cell priming and memory T-cell responses. Although important similarities exist between T cells and NK cells, there are critical differences reflecting their biology and functions. The putative role of PD-1 expression in NK cell immunoregulation has been controversial. Our objective was to comprehensively assess PD-1 expression on NK cells using multiple sources and readouts. Primary human tumor samples, ex vivo culturing, mouse tumors and viral models were all assessed using flow cytometry, qRT-PCR and RNA sequencing. We demonstrate that under multiple activating conditions, highly purified human and mouse NK cells consistently lack PD-1 expression despite the marked upregulation of other regulatory markers such as TIGIT. We further show that neither NK cells from T-cell deficient Rag2-/- mice nor from transgenic PD-1 reporter mice express PD-1 using tumor or viral infection models. Asialo-GM1 (ASGM1), a receptor commonly targeted for NK-specific depletion, was also expressed on activated T cells co-expressing PD-1 contributing to in vivo effects previously attributed to NK cells. These data have important implications when attempting to discern NK from T cell effects depending on the models used and whether PD-1 blockade will directly impact NK cell therapies.
Sean J. Judge, Cordelia Dunai, Ethan G. Aguilar, Sarah C. Vick, Ian R. Sturgill, Lam T. Khuat, Kevin M. Stoffel, Jonathan Van Dyke, Dan L. Longo, Morgan A. Darrow, Stephen K. Anderson, Bruce R. Blazar, Arta M. Monjazeb, Jonathan S. Serody, Robert J. Canter, William J. Murphy
Platinum-based chemotherapy-induced peripheral neuropathy is one of the most common causes of dose reduction and discontinuation of life-saving chemotherapy in cancer treatment; it often causes permanent impairment of quality of life in cancer patients. The mechanisms that underlie this neuropathy are not defined, and effective treatment and prevention measures are not available. Here, we demonstrate that SIRT2 protected mice against cisplatin-induced peripheral neuropathy (CIPN). SIRT2 accumulated in the nuclei of dorsal root ganglion sensory neurons and prevented neuronal cell death following cisplatin treatment. Mechanistically, SIRT2, an NAD+-dependent deacetylase, protected neurons from cisplatin cytotoxicity by promoting transcription-coupled nucleotide excision repair (TC-NER) of cisplatin-induced DNA crosslinks. Consistent with this mechanism, pharmacological inhibition of NER using spironolactone abolished SIRT2-mediated TC-NER activity in differentiated neuronal cells and protection of neurons from cisplatin-induced cytotoxicity and CIPN in mice. Importantly, SIRT2’s protective effects were not evident in lung cancer cells in vitro or in tumors in vivo. Taken together, our results identified SIRT2’s function in the NER pathway as a key underlying mechanism of preventing CIPN, warranting future investigation of SIRT2 activation-mediated neuroprotection during platinum-based cancer treatment.
Manchao Zhang, Wuying Du, Scarlett M. Acklin, Shengkai Jin, Fen Xia
The precise mechanism leading to profound immunodeficiency of HIV-infected patients is still only partially understood. Here, we show that more than 80% of CD4 T cells from HIV-infected patients have morphological abnormalities. Their membranes exhibited numerous large abnormal membrane microdomains (aMMDs), which trap and inactivate physiological receptors, such as that for IL-7. In patient plasma, we identified phospholipase A2 group IB (PLA2G1B) as the key molecule responsible for the formation of aMMDs. At physiological concentrations, PLA2G1B synergized with the HIV gp41 envelope protein, which appears to be a driver that targets PLA2G1B to the CD4 T-cell surface. The PLA2G1B/gp41 pair induced CD4 T cell unresponsiveness (anergy). At high concentrations in vitro, PLA2G1B acted alone, independently of gp41, and inhibited the IL-2, IL-4, and IL-7 responses, as well as TCR-mediated activation and proliferation, of CD4 T cells. PLA2G1B also decreased CD4 T-cell survival in vitro, likely playing a role in CD4 lymphopenia in conjunction with its induced IL-7 receptor defects. The effects on CD4 T-cell anergy could be blocked by a PLA2G1B-specific neutralizing mAb in vitro and in vivo. The PLA2G1B/gp41 pair constitutes a new mechanism of immune dysfunction and a compelling target for boosting immune responses in HIV-infected patients.
Julien Pothlichet, Thierry Rose, Florence Bugault, Louise Jeammet, Annalisa Meola, Ahmed Haouz, Frederick Saul, David Geny, José Alcami, Ezequiel Ruiz-Mateos Carmona, Luc Teyton, Gérard Lambeau, Jacques Thèze