Expanded Histopathology and Tropism of Ebola Virus in the Rhesus Macaque Model: Potential for Sexual Transmission, Altered Adrenomedullary Hormone Production and Early Viral Replication in liver

The American journal of pathology

2021 Oct 06

Liu, DX;Cooper, TK;Perry, DL;Huzella, LM;Hischak, AM;Hart, RJ;Isic, N;Byrum, R;Ragland, D;St Claire, M;Cooper, K;Reeder, R;Logue, J;Jahrling, PB;Holbrook, MR;Bennett, RS;Hensley, LE;
PMID: 34626576 | DOI: 10.1016/j.ajpath.2021.09.009

The pathogenesis of Ebola virus disease (EVD) is still incomplete, although the non-human primate model has been studied for more than 4 decades. To further investigate EVD pathogenesis, a natural history study has been conducted using 27 Chinese-origin rhesus macaques. Of them, 24 macaques were exposed intramuscularly to Kikwit Ebola virus (EBOV) and euthanized at pre-determined timepoints or when end stage clinical disease criteria were met, while 3 other sham-exposed macaques were euthanized at the study day 0. This study demonstrates for the first time that Ebola virus causes uterine cervicitis, vaginitis, posthitis, and medullary adrenalitis. Not only is Ebola virus detected in the interstitial stromal cells of the genital tract, but it is also present in the epididymal and seminal vesicular tubular epithelial cells, ectocervical and vaginal squamous epithelial cells, and seminal fluid. Furthermore, as early as day 3 after exposure, EBOV replicative intermediate RNA was detected in Kupffer cells and hepatocytes. These findings in the nonhuman model provide additional insight into potential sexual transmission, possible disruption of sympathetic hormone production, and early virus replication sites in human EVD patients.

Detection of rabbit hemorrhagic disease virus 2 in formalin-fixed, paraffin-embedded tissues via in situ hybridization

Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc

2021 Sep 23

O'Toole, AD;Zhang, J;Williams, LBA;Brown, CC;
PMID: 34554024 | DOI: 10.1177/10406387211047561

Formalin-fixed, paraffin-embedded tissues from European rabbits (Oryctolagus cuniculus) that succumbed to rabbit hemorrhagic disease virus 2 (RHDV2; Lagovirus GI.2) during the 2019 outbreak in Washington, USA, were utilized for in situ hybridization via RNAscope (ACDBio). This detection method was both sensitive and specific, with no staining in tissues from RHDV- (Lagovirus GI.1) and RHDV2-negative rabbits, and only slight background staining of RHDV-positive rabbits; RHDV2-positive tissues had bright-red cytoplasmic staining. Although much of the viral mRNA detection was consistent with previously described antigen detection via immunohistochemistry of the liver, lungs, and spleen, there was also significant glomerular staining in the kidneys, and endothelial staining within blood vessels of almost all organs. We validated the RNAscope technique for detection of RHDV2 mRNA in formalin-fixed, paraffin-embedded tissues, with increased sensitivity from previous techniques, and identified additional affected cell types that may contribute to the understanding of pathogenesis.

An RNA aptamer restores defective bone growth in FGFR3-related skeletal dysplasia in mice

Science translational medicine

2021 May 05

Kimura, T;Bosakova, M;Nonaka, Y;Hruba, E;Yasuda, K;Futakawa, S;Kubota, T;Fafilek, B;Gregor, T;Abraham, SP;Gomolkova, R;Belaskova, S;Pesl, M;Csukasi, F;Duran, I;Fujiwara, M;Kavkova, M;Zikmund, T;Kaiser, J;Buchtova, M;Krakow, D;Nakamura, Y;Ozono, K;Krejci, P;
PMID: 33952673 | DOI: 10.1126/scitranslmed.aba4226

Achondroplasia is the most prevalent genetic form of dwarfism in humans and is caused by activating mutations in FGFR3 tyrosine kinase. The clinical need for a safe and effective inhibitor of FGFR3 is unmet, leaving achondroplasia currently incurable. Here, we evaluated RBM-007, an RNA aptamer previously developed to neutralize the FGFR3 ligand FGF2, for its activity against FGFR3. In cultured rat chondrocytes or mouse embryonal tibia organ culture, RBM-007 rescued the proliferation arrest, degradation of cartilaginous extracellular matrix, premature senescence, and impaired hypertrophic differentiation induced by FGFR3 signaling. In cartilage xenografts derived from induced pluripotent stem cells from individuals with achondroplasia, RBM-007 rescued impaired chondrocyte differentiation and maturation. When delivered by subcutaneous injection, RBM-007 restored defective skeletal growth in a mouse model of achondroplasia. We thus demonstrate a ligand-trap concept of targeting the cartilage FGFR3 and delineate a potential therapeutic approach for achondroplasia and other FGFR3-related skeletal dysplasias.

A role of PIEZO1 in iron metabolism in mice and humans

Cell

2021 Feb 18

Ma, S;Dubin, AE;Zhang, Y;Mousavi, SAR;Wang, Y;Coombs, AM;Loud, M;Andolfo, I;Patapoutian, A;
PMID: 33571427 | DOI: 10.1016/j.cell.2021.01.024

Iron overload causes progressive organ damage and is associated with arthritis, liver damage, and heart failure. Elevated iron levels are present in 1%-5% of individuals; however, iron overload is undermonitored and underdiagnosed. Genetic factors affecting iron homeostasis are emerging. Individuals with hereditary xerocytosis, a rare disorder with gain-of-function (GOF) mutations in mechanosensitive PIEZO1 ion channel, develop age-onset iron overload. We show that constitutive or macrophage expression of a GOF Piezo1 allele in mice disrupts levels of the iron regulator hepcidin and causes iron overload. We further show that PIEZO1 is a key regulator of macrophage phagocytic activity and subsequent erythrocyte turnover. Strikingly, we find that E756del, a mild GOF PIEZO1 allele present in one-third of individuals of African descent, is strongly associated with increased plasma iron. Our study links macrophage mechanotransduction to iron metabolism and identifies a genetic risk factor for increased iron levels in African Americans.

AT2 cell-derived IgA trapped by the extracellular matrix in silica-induced pulmonary fibrosis

International immunopharmacology

2023 Jun 28

Chen, M;Wang, J;Yuan, M;Long, M;Sun, Y;Wang, S;Luo, W;Zhou, Y;Zhang, W;Jiang, W;Chao, J;
PMID: 37390644 | DOI: 10.1016/j.intimp.2023.110545

Pulmonary fibrosis is an interstitial lung disease caused by various factors such as exposure to workplace environmental contaminants, drugs, or X-rays. Epithelial cells are among the driving factors of pulmonary fibrosis. Immunoglobulin A (IgA), traditionally thought to be secreted by B cells, is an important immune factor involved in respiratory mucosal immunity. In the current study, we found that lung epithelial cells are involved in IgA secretion, which, in turn, promotes pulmonary fibrosis. Spatial transcriptomics and single-cell sequencing suggest that Igha transcripts were highly expressed in the fibrotic lesion areas of lungs from silica-treated mice. Reconstruction of B-cell receptor (BCR) sequences revealed a new cluster of AT2-like epithelial cells with a shared BCR and high expression of genes related to IgA production. Furthermore, the secretion of IgA by AT2-like cells was trapped by the extracellular matrix and aggravated pulmonary fibrosis by activating fibroblasts. Targeted blockade of IgA secretion by pulmonary epithelial cells may be a potential strategy for treating pulmonary fibrosis.

MHC class II-restricted antigen presentation is required to prevent dysfunction of cytotoxic T cells by blood-borne myeloids in brain tumors

Cancer cell

2022 Dec 30

Kilian, M;Sheinin, R;Tan, CL;Friedrich, M;Krämer, C;Kaminitz, A;Sanghvi, K;Lindner, K;Chih, YC;Cichon, F;Richter, B;Jung, S;Jähne, K;Ratliff, M;Prins, RM;Etminan, N;von Deimling, A;Wick, W;Madi, A;Bunse, L;Platten, M;
PMID: 36638785 | DOI: 10.1016/j.ccell.2022.12.007

Cancer immunotherapy critically depends on fitness of cytotoxic and helper T cell responses. Dysfunctional cytotoxic T cell states in the tumor microenvironment (TME) are a major cause of resistance to immunotherapy. Intratumoral myeloid cells, particularly blood-borne myeloids (bbm), are key drivers of T cell dysfunction in the TME. We show here that major histocompatibility complex class II (MHCII)-restricted antigen presentation on bbm is essential to control the growth of brain tumors. Loss of MHCII on bbm drives dysfunctional intratumoral tumor-reactive CD8+ T cell states through increased chromatin accessibility and expression of Tox, a critical regulator of T cell exhaustion. Mechanistically, MHCII-dependent activation of CD4+ T cells restricts myeloid-derived osteopontin that triggers a chronic activation of NFAT2 in tumor-reactive CD8+ T cells. In summary, we provide evidence that MHCII-restricted antigen presentation on bbm is a key mechanism to directly maintain functional cytotoxic T cell states in brain tumors.

LncRNA LIMp27 Regulates the DNA Damage Response through p27 in p53-Defective Cancer Cells

Advanced science (Weinheim, Baden-Wurttemberg, Germany)

2023 Jan 13

La, T;Chen, S;Zhao, XH;Zhou, S;Xu, R;Teng, L;Zhang, YY;Ye, K;Xu, L;Guo, T;Jamaluddin, MF;Feng, YC;Tang, HJ;Wang, Y;Xu, Q;Gu, Y;Cao, H;Liu, T;Thorne, RF;Shao, FM;Zhang, XD;Jin, L;
PMID: 36638271 | DOI: 10.1002/advs.202204599

P53 inactivation occurs in about 50% of human cancers, where p53-driven p21 activity is devoid and p27 becomes essential for the establishment of the G1/S checkpoint upon DNA damage. Here, this work shows that the E2F1-responsive lncRNA LIMp27 selectively represses p27 expression and contributes to proliferation, tumorigenicity, and treatment resistance in p53-defective colon adenocarcinoma (COAD) cells. LIMp27 competes with p27 mRNA for binding to cytoplasmically localized hnRNA0, which otherwise stabilizes p27 mRNA leading to cell cycle arrest at the G0/G1 phase. In response to DNA damage, LIMp27 is upregulated in both wild-type and p53-mutant COAD cells, whereas cytoplasmic hnRNPA0 is only increased in p53-mutant COAD cells due to translocation from the nucleus. Moreover, high LIMp27 expression is associated with poor survival of p53-mutant but not wild-type p53 COAD patients. These results uncover an lncRNA mechanism that promotes p53-defective cancer pathogenesis and suggest that LIMp27 may constitute a target for the treatment of such cancers.

Hypothalamic warm-sensitive neurons require TRPC4 channel for detecting internal warmth and regulating body temperature in mice

Neuron

2022 Nov 29

Zhou, Q;Fu, X;Xu, J;Dong, S;Liu, C;Cheng, D;Gao, C;Huang, M;Liu, Z;Ni, X;Hua, R;Tu, H;Sun, H;Shen, Q;Chen, B;Zhang, J;Zhang, L;Yang, H;Hu, J;Yang, W;Pei, W;Yao, Q;Sheng, X;Zhang, J;Yang, WZ;Shen, WL;
PMID: 36476978 | DOI: 10.1016/j.neuron.2022.11.008

Precise monitoring of internal temperature is vital for thermal homeostasis in mammals. For decades, warm-sensitive neurons (WSNs) within the preoptic area (POA) were thought to sense internal warmth, using this information as feedback to regulate body temperature (Tcore). However, the cellular and molecular mechanisms by which WSNs measure temperature remain largely undefined. Via a pilot genetic screen, we found that silencing the TRPC4 channel in mice substantially attenuated hypothermia induced by light-mediated heating of the POA. Loss-of-function studies of TRPC4 confirmed its role in warm sensing in GABAergic WSNs, causing additional defects in basal temperature setting, warm defense, and fever responses. Furthermore, TRPC4 antagonists and agonists bidirectionally regulated Tcore. Thus, our data indicate that TRPC4 is essential for sensing internal warmth and that TRPC4-expressing GABAergic WSNs function as a novel cellular sensor for preventing Tcore from exceeding set-point temperatures. TRPC4 may represent a potential therapeutic target for managing Tcore.

Association of sarcoidosis with psoriasis: a cross-sectional study in the All of Us research program

Archives of dermatological research

2022 Nov 27

Murphy, MJ;Leasure, AC;Damsky, W;Cohen, JM;
PMID: 36436011 | DOI: 10.1007/s00403-022-02488-z

Psoriasis and sarcoidosis are inflammatory skin and systemic diseases that may share a similar immunopathogenesis involving a Th1 and/or Th17 polarized immune response. Although the coexistence of sarcoidosis and psoriasis in the same individuals has been reported, the potential association between these diseases at a population-level in the United States has not been evaluated. To evaluate this association, we performed a matched cross-sectional study in the All of Us research program database. In the multivariable analysis of 4932 psoriasis cases and 19,728 controls, sarcoidosis was found to be significantly associated with psoriasis (OR 2.37 [95% CI 1.73-3.23], p < 0.001). The relative strength of this association between psoriasis and sarcoidosis may be, in part, explained by overlapping immunopathogenesis and common genetic susceptibility of these diseases. Taken together, these observations underscore the need for screening psoriasis patients for development of new cardiopulmonary symptoms. Further research into the mechanism of this relationship and its implications is warranted.

HnRNP K mislocalisation in neurons of the dentate nucleus is a novel neuropathological feature of neurodegenerative disease and ageing

Neuropathology and applied neurobiology

2022 Jun 01

Sidhu, R;Gatt, A;Fratta, P;Lashley, T;Bampton, A;
PMID: 35064577 | DOI: 10.1111/nan.12793

Nuclear depletion and cytoplasmic mislocalisation of the RNA-binding protein heterogeneous ribonucleoprotein K (hnRNP K) within pyramidal neurons of the frontal cortex have been shown to be a common neuropathological feature in frontotemporal lobar degeneration (FTLD) and elderly control brain. Here, we describe a second neuronal subtype vulnerable to mislocalisation within the dentate nucleus of the cerebellum. In contrast to neurons within the cerebellar cortex that typically exhibited normal, nuclear staining, many neurons of the dentate nucleus exhibited striking mislocalisation of hnRNP K to the cytoplasm within neurodegenerative disease brain. Mislocalisation frequency in this region was found to be significantly higher in both FTLD-TDP A and Alzheimer's disease (AD) brain than in age-matched controls. However, within control (but not disease) subjects, mislocalisation frequency was significantly associated with age-at-death with more elderly controls typically exhibiting greater levels of the pathology. This study provides further evidence for hnRNP K mislocalisation being a more anatomically diverse pathology than previously thought and suggests that potential dysfunction of the protein may be more broadly relevant to the fields of neurodegeneration and ageing.

Profiling cell-type specific gene expression in post-mortem human brain samples through laser capture microdissection

Methods (San Diego, Calif.)

2022 Sep 03

Almeida, D;Turecki, G;
PMID: 36064002 | DOI: 10.1016/j.ymeth.2022.08.013

The transcriptome of a cell constitutes an essential piece of cellular identity and contributes to the multifaceted complexity and heterogeneity of cell-types within the mammalian brain. Thus, while a wealth of studies have investigated transcriptomic alterations underlying the pathophysiology of diseases of the brain, their use of bulk-tissue homogenates makes it difficult to tease apart whether observed differences are explained by disease state or cellular composition. Cell-type-specific enrichment strategies are, therefore, crucial in the context of gene expression profiling. Laser capture microdissection (LCM) is one such strategy that allows for the capture of specific cell-types, or regions of interest, under microscopic visualization. In this review, we focus on using LCM for cell-type specific gene expression profiling in post-mortem human brain samples. We begin with a discussion of various LCM systems, followed by a walk-through of each step in the LCM to gene expression profiling workflow and a description of some of the limitations associated with LCM. Throughout the review, we highlight important considerations when using LCM with post-mortem human brain samples. Whenever applicable, commercially available kits that have proven successful in the context of LCM with post-mortem human brain samples are described.

Light receptors in the avian brain and seasonal reproduction

Journal of experimental zoology. Part A, Ecological and integrative physiology

2022 Sep 02

Pérez, JH;
PMID: 36052512 | DOI: 10.1002/jez.2652

Detection and transduction of photic cues by nonvisual photoreceptors, located in the deep brain, is a critical component of timing seasonal reproduction in birds. However, the precise identity of the photoreceptors responsible for detection of salient photic cues remains uncertain and debated. Here I review of the existing evidence for each of the three candidate photoreceptive opsins: Vertebrate Ancient Opsin, Melanopsin, and Neuropsin, including localization, action spectrum, and data from experimental manipulation of opsin expression. These findings are compared to an updated list of key criteria established in the literature as a litmus for classifying an opsin as the "breeding photoreceptor." Integrating evidence for each of the candidate photoreceptors with respect to these criteria reveals support for all three opsins in regulation of seasonal reproduction. Taken together these findings strongly suggest that transduction of seasonal photoperiodic information involves the activity of multiple photoreceptor types and populations functioning in concert. This review also highlights the need to shift attention from simply identifying "the breeding photoreceptor" to a more integrative approach aiming to parse the contribution of specific photoreceptor populations within the brain.

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	Description
sense Example: Hs-LAG3-sense	Standard probes for RNA detection are in antisense. Sense probe is reverse complent to the corresponding antisense probe.
Intron# Example: Mm-Htt-intron2	Probe targets the indicated intron in the target gene, commonly used for pre-mRNA detection
Pool/Pan Example: Hs-CD3-pool (Hs-CD3D, Hs-CD3E, Hs-CD3G)	A mixture of multiple probe sets targeting multiple genes or transcripts
No-XSp Example: Hs-PDGFB-No-XMm	Does not cross detect with the species (Sp)
XSp Example: Rn-Pde9a-XMm	designed to cross detect with the species (Sp)
O# Example: Mm-Islr-O1	Alternative design targeting different regions of the same transcript or isoforms
CDS Example: Hs-SLC31A-CDS	Probe targets the protein-coding sequence only
EnEm	Probe targets exons n and m
En-Em	Probe targets region from exon n to exon m
Retired Nomenclature
tvn Example: Hs-LEPR-tv1	Designed to target transcript variant n
ORF Example: Hs-ACVRL1-ORF	Probe targets open reading frame
UTR Example: Hs-HTT-UTR-C3	Probe targets the untranslated region (non-protein-coding region) only
5UTR Example: Hs-GNRHR-5UTR	Probe targets the 5' untranslated region only
3UTR Example: Rn-Npy1r-3UTR	Probe targets the 3' untranslated region only
Pan Example: Pool	A mixture of multiple probe sets targeting multiple genes or transcripts

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