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Abstract
BACKGROUND. Prostate cancer is multifocal with distinct molecular subtypes. The utility of genomic subtyping has been challenged due to inter- and intra-focal heterogeneity. We sought to characterize the subtype-defining molecular alterations of primary prostate cancer across all tumor foci within radical prostatectomy (RP) specimens and determine the prevalence of collision tumors. METHODS. From the Early Detection Research Network cohort, we identified 333 prospectively collected RPs from 2010 to 2014 and assessed ERG, SPINK1, PTEN, and SPOP molecular status. We utilized dual ERG/SPINK1 immunohistochemistry, fluorescence in situ hybridization to confirm ERG rearrangements and characterize PTEN deletion, and high-resolution melting curve analysis and Sanger sequencing to determine SPOP mutation status. Analysis of biochemical recurrence-free of patients with collision tumors was conducted using Kaplan-Meier method. RESULTS. Based on index focus alone, ERG, SPINK1, PTEN, and SPOP alterations were identified in 47.5%, 10.8%, 14.3%, and 5.1% of RP specimens, respectively. In 233 multifocal RPs with ERG/SPINK1 status in all foci, 139 (59.7%) had discordant molecular alterations between foci. Collision tumors, as defined by discrepant ERG/SPINK1 status within a single focus, were identified in 29 (9.4%) RP specimens. CONCLUSION. Interfocal molecular heterogeneity was identified in ~60% of multifocal RP specimens and collision tumors were present in ~10%. We present this phenomenon as a model for the intra-focal heterogeneity observed in previous studies and propose future genomic studies screen for collision tumors to better characterize molecular heterogeneity.
Authors
Jacqueline Fontugne, Peter Y. Cai, Hussein Alnajar, Bhavneet Bhinder, Kyung Park, Huihui Ye, Shaham Beg, Verena Sailer, Javed Siddiqui, Mirjam Blattner-Johnson, Jaclyn A. Croyle, Zohal Noorzad, Carla Calagua, Theresa Y. MacDonald, Ulrika Axcrona, Mari Bogaard, Karol Axcrona, Douglas S. Scherr, Martin G. Sanda, Bjarne Johannessen, Arul M. Chinnaiyan, Olivier Elemento, Rolf I. Skotheim, Mark A. Rubin, Christopher E. Barbieri, Juan M. Mosquera
Abstract
Why Multisystem Inflammatory Syndrome in Children (MIS-C) develops after SARS-CoV-2 infection in a subset of children is unknown. We hypothesized that aberrant virus52 specific T-cell responses contribute to MIS-C pathogenesis. We quantified SARS-CoV-2 reactive T-cells, serologic responses against major viral proteins, and cytokine responses from plasma and peripheral blood mononuclear cells in children with convalescent COVID-19, acute MIS-C, and healthy controls. Children with MIS-C had significantly lower virus-specific CD4+ and CD8+ T-cell responses to major SARS-CoV-2 antigens compared with children convalescing from COVID-19. Further, T-cell responses in participants with MIS-C were similar to or lower than those in healthy controls. Serologic responses against spike receptor binding domain (RBD), full-length spike, and nucleocapsid were similar among convalescent COVID-19 and MIS-C, suggesting functional B cell responses. Cytokine profiling demonstrated predominant Th1 polarization of CD4+ T-cells from children with convalescent COVID-19 and MIS-C, although cytokine production was reduced in MIS-C. Our findings support a role for constrained induction of anti-SARS-CoV-2-specific T-cells in the pathogenesis of MIS-C.
Authors
Vidisha Singh, Veronica Obregon-Perko, Stacey A. Lapp, Anna M. Horner, Alyssa Brooks, Lisa Macoy, Laila Hussaini, Austin Lu, Theda Gibson, Guido Silvestri, Alba Grifoni, Daniela Weiskopf, Alessandro Sette, Evan J. Anderson, Christina A. Rostad, Ann Chahroudi
Abstract
Impaired glucose metabolism is observed in obesity and type 2 diabetes. Glucose controls gene expression through the transcription factor ChREBP in liver and adipose tissues. Mlxipl encodes two isoforms, ChREBPα, the full-length form which translocation into the nucleus is under the control of glucose and, ChREBPβ, a constitutively nuclear shorter form. ChREBPβ gene expression in white adipose tissue is strongly associated with insulin sensitivity. Here, we investigated the consequences of ChREBPβ deficiency on insulin action and energy balance. ChREBPβ-deficient male and female C57BL6/J and FVB/N mice were produced using CRISPR-Cas9-mediated gene editing. Unlike global ChREBP deficiency, lack of ChREBPβ showed modest effects on gene expression in adipose tissues and liver, with variations seen chiefly in brown adipose tissue. In mice fed chow and high fat diets, lack of ChREBPβ had moderate effects on body composition and insulin sensitivity. ChREBPβ deficiency did not prevent the whitening of brown adipose tissue reported in total ChREBP knock out mice at thermoneutrality. These findings reveal that ChREBPβ is dispensable for metabolic adaptations to nutritional and thermic challenges.
Authors
Emeline Recazens, Geneviève Tavernier, Jérémy Dufau, Camille Bergoglio, Fadila Benhamed, Stéphanie Cassant-Sourdy, Marie-Adeline Marques, Sylvie Caspar-Bauguil, Alice Brion, Laurent Monbrun, Renaud Dentin, Clara Ferrier, Mélanie Leroux, Pierre-Damien Denechaud, Cedric Moro, Jean-Paul Concordet, Catherine Postic, Etienne Mouisel, Dominique Langin
Abstract
Total body irradiation (TBI) targets sensitive bone marrow hematopoietic cells and gut epithelial cells causing their death and TBI induces the state of immunodeficiency combined with intestinal dysbiosis and non-productive immune responses. We found enhanced Pseudomonas aeruginosa (PA) colonization of the gut leading to the host cell death and strikingly decreased survival of irradiated mice. PA-driven pathogenic mechanism includes theft-ferroptosis is realized via: i) curbing host anti-ferroptotic system GSH/GPx4 and ii) employing bacterial 15-lipoxygenase (pLoxA) to generate pro-ferroptotic signal - 15-hyderoperoxy-arachidonyl-PE (15-HpETE-PE) - in the intestines of irradiated/infected mice. Global redox phospholipidomics of the ileum revealed that lyso-phospholipids and oxidized phospholipids (particularly PEox) represented the major factors which contributed to the TBI+PA induced pathogenic changes. A lipoxygenase inhibitor, baicalein, significantly attenuated animal lethality, PA colonization, as well as intestinal epithelial cell death and generation of ferroptotic PEox signals. Opportunistic PA mechanisms included stimulation of the anti-inflammatory lipoxin A4 (LXA4) production and suppression of the pro-inflammatory hepoxilin A3 (HxA3) and leukotriene B4 (LTB4). Unearthing complex PA pathogenic/virulence mechanisms including effects on the host anti-/pro-inflammatory responses, lipid metabolism and ferroptotic cell death points to new therapeutic and radiomitigative targets.
Authors
Haider H. Dar, Michael W. Epperly, Vladimir A. Tyurin, Andrew A. Amoscato, Tamil S. Anthonymuthu, Austin B. Souryavong, Alexander A. Kapralov, Galina V. Shurin, Svetlana N. Samovich, Claudette M. St. Croix, Simon C. Watkins, Sally E. Wenzel, Rama K. Mallampalli, Joel S. Greenberger, Hulya Bayir, Valerian E. Kagan, Yulia Y. Tyurina
Abstract
Defective primary cilia cause a range of diseases called ciliopathies, which include hearing loss (HL). Variants in human oxysterol binding protein like 2 (OSBPL2/ORP2) are responsible for autosomal dominant nonsyndromic HL (DFNA67). However, the pathogenesis of OSBPL2 deficiency has not been fully elucidated. In this study, we showed that the Osbpl2-knockout (KO) mice exhibited progressive HL and abnormal cochlea development with defective cilia. Further research revealed that OSBPL2 was located at the base of kinocilia in hair cells (HCs) and primary cilia in supporting cells (SCs), and functioned in the maintenance of ciliogenesis by regulating the homeostasis of PI(4,5)P2 on the cilia membrane. OSBPL2 deficiency led to a significant increase of PI(4,5)P2 on the cilia membrane, which could be partially rescued by the overexpression of INPP5E. In addition, the key molecules in Sonic Hedgehog (Shh) signaling pathway (SMO and GLI3) were detected to be down-regulated in Osbpl2-KO HEI-OC1 cells. Our findings revealed that OSBPL2 deficiency resulted in ciliary defects and abnormal Shh signaling transduction in auditory cells, which helped to elucidate the underlying mechanism of OSBPL2 deficiency in HL.
Authors
Hairong Shi, Hongshun Wang, Cheng Zhang, Yajie Lu, Jun Yao, Zhibin Chen, Guangqian Xing, Qinjun Wei, Xin Cao
Abstract
BACKGROUND. Vaccine-elicited adaptive immunity is a prerequisite for control of SARS-CoV-2 infection. Multiple sclerosis (MS) disease-modifying therapies (DMTs) differentially target humoral and cellular immunity. A comprehensive comparison of MS DMTs on SARS-CoV-2 vaccine-specific immunity is needed, including quantitative and functional B and T cell responses. METHODS. Spike-specific antibody and T cell responses were measured before and following SARS-CoV-2 vaccination in a cohort of 80 subjects, including healthy controls and MS patients in six DMT groups: untreated, glatiramer acetate (GA), dimethyl fumarate (DMF), natalizumab (NTZ), sphingosine-1-phosphate (S1P) receptor modulators, and anti-CD20 monoclonal antibodies. Anti-spike antibody responses were quantified by Luminex assay, high-resolution spike epitope reactivity was mapped by VirScan, and pseudovirus neutralization was assessed. Spike-specific CD4+ and CD8+ T cell responses were characterized by activation-induced marker (AIM) expression, cytokine production, and tetramer analysis. RESULTS. Anti-spike IgG levels were similar between healthy controls, untreated MS, GA, DMF, and NTZ patients, but were significantly reduced in anti-CD20 and S1P-treated patients. Anti-spike seropositivity in anti-CD20 patients was significantly correlated with CD19+ B cell levels and inversely correlated with cumulative treatment duration. Spike epitope reactivity and pseudovirus neutralization was reduced in anti-CD20 and S1P patients, directly correlating with reduced spike receptor binding domain (RBD) IgG levels. Spike-specific CD4+ and CD8+ T cell reactivity remained robust across all groups except in S1P-treated patients in whom post-vaccine CD4+ T cell responses were attenuated. CONCLUSIONS. These findings from a large MS cohort exposed to a wide spectrum of MS immunotherapies have important implications for treatment-specific COVID-19 clinical guidelines. FUNDING. This work was supported by grants from the NIH 1K08NS107619 (JJS), NMSS TA- 1903-33713 (JJS), K08NS096117 (MRW), Westridge Foundation (MRW), Chan Zuckerberg Biohub (JLD), R01AI159260 (JAH), R01NS092835 (SSZ), R01AI131624 (SSZ), R21NS108159 (SSZ), NMSS RG1701-26628 (SSZ), and the Maisin Foundation (SSZ).
Authors
Joseph J. Sabatino Jr, Kristen Mittl, William M. Rowles, Kira McPolin, Jayant V. Rajan, Matthew T. Laurie, Colin R. Zamecnik, Ravi Dandekar, Bonny D. Alvarenga, Rita P. Loudermilk, Chloe Gerungan, Collin M. Spencer, Sharon A. Sagan, Danillo G. Augusto, Jessa R. Alexander, Joseph L. DeRisi, Jill A. Hollenbach, Michael R. Wilson, Scott S. Zamvil, Riley Bove
Abstract
Aristolochic acid (AA) is the causative nephrotoxic alkaloid in aristolochic acid nephropathy, which results in a tubulointerstitial fibrosis. AA causes direct proximal tubule damage. There is also an influx of macrophages, although their role in the pathogenesis is poorly understood. Here we demonstrate that AA directly stimulates migration, inflammation, and reactive oxygen species (ROS) production in macrophages ex vivo. Cells lacking interferon regulatory factor 4 (IRF4), a known regulator of macrophage migration and phenotype, had a reduced migratory response, though effects on ROS production and inflammation were preserved or increased relative to wild-type cells. Macrophage-specific IRF4 knockout mice were protected from both acute and chronic kidney effects of AA administration based on functional and histological analysis. Renal macrophages from kidneys of AA-treated macrophage-specific IRF4 knockout mice demonstrated increased apoptosis and ROS production compared with wildtype controls, indicating that AA directly polarizes macrophages to a promigratory and proinflammatory phenotype. However, knockout mice had reduced renal macrophage abundance following AA administration. While macrophages lacking IRF4 can adopt a proinflammatory phenotype upon AA exposure, their inability to migrate to the kidney and increased rates of apoptosis upon infiltration provide protection from AA in vivo. These results provide evidence of direct AA effects on macrophages in AAN and add to the growing body of evidence that supports a key role of IRF4 in modulating macrophage function in kidney injury.
Authors
Kensuke Sasaki, Andrew S. Terker, Jiaqi Tang, Shirong Cao, Juan Pablo Arroyo, Aolei Niu, Suwan Wang, Xiaofeng Fan, Yahua Zhang, Stephanie R. Bennett, Ming-zhi Zhang, Raymond C. Harris
Abstract
Mammalian skeletal muscle contains heterogenous myofibers with different contractile and metabolic properties that sustain muscle mass and endurance capacity. The transcriptional regulators that govern these myofiber gene programs have been elucidated. However, the hormonal cues that direct the specification of myofiber types and muscle endurance remain largely unknown. Here we uncover the secreted factor Tsukushi (TSK) as an extracellular signal that is required for maintaining muscle mass, strength, and endurance capacity, and contributes to muscle regeneration. Mice lacking TSK exhibited reduced grip strength and impaired exercise capacity. Muscle transcriptomic analysis revealed that TSK deficiency results in a remarkably selective impairment in the expression of myofibrillar genes characteristic of slow-twitch muscle fibers that is associated with abnormal neuromuscular junction formation. AAV-mediated overexpression of TSK failed to rescue these myofiber defects in adult mice, suggesting that the effects of TSK on myofibers are likely restricted to certain developmental stages. Finally, mice lacking TSK exhibited diminished muscle regeneration following cardiotoxin-induced muscle injury. These findings support a crucial role of TSK as a hormonal cue in the regulation of contractile gene expression, endurance capacity, and muscle regeneration.
Authors
Qiuyu Wang, Xiaoxue Qiu, Tongyu Liu, Cheehoon Ahn, Jeffrey F. Horowitz, Jiandie D. Lin
Abstract
Recent research on altering threat memory has focused on a reconsolidation window. During reconsolidation, threat memories are retrieved and become labile. Reconsolidation of distinct threat memories is synapse-dependent whereas the underlying regulatory mechanism of the specificity of reconsolidation is poorly understood. We designed a unique behavioral paradigm in which a distinct threat memory can be retrieved through the associated conditioned stimulus. In addition, we proposed a regulatory mechanism by which the activation of acid-sensing ion channels (ASICs), strengthens the distinct memory trace associated with the memory reconsolidation to determine its specificity. The activation of ASICs by carbon dioxide (CO2) inhalation when paired with memory retrieval, triggers the reactivation of the distinct memory trace, resulting in greater memory lability. ASICs potentiate the memory trace by altering the amygdala-dependent synaptic transmission and plasticity at selectively targeted synapses. Our results suggest that inhaling CO2 during the retrieval event increases the lability of a threat memory through a synapse-specific reconsolidation process.
Authors
Erin E. Koffman, Charles M. Kruse, Kritika Singh, Farzaneh Sadat Naghavi, Melissa A. Curtis, Jennifer Egbo, Mark Houdi, Boren Lin, Hui Lu, Jacek Debiec, Jianyang Du
Abstract
BACKGROUND. Most subjects with prior COVID-19 disease manifest long–term, protective immune responses against re-infection. Accordingly, we tested the hypothesis that humoral immune and reactogenicity responses to a SARS-CoV-2 mRNA vaccine differ in subjects with and without prior COVID-19. METHODS. Health care workers (n=61) with (n=30) and without (n=31) prior COVID-19 disease received two, 30 µg doses of Pfizer BNT162b2 vaccine 3 weeks apart. Serum IgG antibody against the Spike receptor-binding domain (RBD); serum neutralizing activity; and vaccine reactogenicity were assessed longitudinally every 2 weeks for 56 days after the 1st injection. RESULTS. The COVID group manifested more rapid increases in Spike IgG antibody and serum neutralizing activity post 1st vaccine dose but showed little or no increase after the 2nd dose compared to the infection-naïve group. In fact, Spike IgG was maximum after the 1st dose in 36% of the COVID group versus 0% of the infection-naïve group. Peak IgG antibody was lower but appeared to fall more slowly in the COVID-19 versus the infection-naïve group. Finally, adverse systemic reactions e.g., fever, headache and malaise, were more frequent and lasted longer after both the 1st and 2nd injection in the COVID group than in the infection-naïve group. CONCLUSION. Subjects with prior COVID-19 demonstrate a robust, accelerated humoral immune response to the 1st dose but attenuated response to the 2nd dose of BNT162b2 vaccine compared to controls. The COVID-19 group also experiences greater reactogenicity. Humoral responses and reactogenicity to BNT162b2 differ qualitatively and quantitatively in subjects with prior COVID-19 compared to infection-naive subjects. FUNDING. This work was supported by Institutional Funds.
Authors
Steven G. Kelsen, Alan S. Braverman, Mark O. Aksoy, Jacob A. Hayman, Puja S. Patel, Charu Rajput, Huaqing Zhao, Susan G. Fisher, Michael R. Ruggieri Sr., Nina T. Gentile
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