Activation of STAT3 signaling pathway in the kidney of COVID-19 patients

Journal of nephrology

2021 Oct 09

Salem, F;Li, XZ;Hindi, J;Casablanca, NM;Zhong, F;El Jamal, SM;Haroon Al Rasheed, MR;Li, L;Lee, K;Chan, L;He, JC;
PMID: 34626364 | DOI: 10.1007/s40620-021-01173-0

Acute kidney injury is common in patients with COVID-19, however mechanisms of kidney injury remain unclear. Since cytokine storm is likely a cause of AKI and glomerular disease, we investigated the two major transcription factors, STAT3 and NF-kB, which are known to be activated by cytokines.This is an observational study of the postmortem kidneys of 50 patients who died with COVID-19 in the Mount Sinai Hospital during the first pandemic surge. All samples were reviewed under light microscopy, electron microscopy, and immunofluorescence by trained renal pathologists. In situ hybridization evaluation for SARS-CoV-2 and immunostaining of transcription factors STAT3 and NF-kB were performed.Consistent with previous findings, acute tubular injury was the major pathological finding, together with global or focal glomerulosclerosis. We were not able to detect SARS-CoV-2 in kidney cells. ACE2 expression was reduced in the tubular cells of patients who died with COVID-19 and did not co-localize with TMPRSS2. SARS-CoV-2 was identified occasionally in the mononuclear cells in the peritubular capillary and interstitium. STAT3 phosphorylation at Tyr705 was increased in 2 cases in the glomeruli and in 3 cases in the tubulointerstitial compartments. Interestingly, STAT3 phosphorylation at Ser727 increased in 9 cases but only in the tubulointerstitial compartment. A significant increase in NF-kB phosphorylation at Ser276 was also found in the tubulointerstitium of the two patients with increased p-STAT3 (Tyr705).Our findings suggest that, instead of tyrosine phosphorylation, serine phosphorylation of STAT3 is commonly activated in the kidney of patients with COVID-19.

Evidence of vertical transmission of SARS-CoV-2 and interstitial pneumonia in second trimester twin stillbirth in asymptomatic woman. Case report and review of the literature

American journal of obstetrics & gynecology MFM

2022 Feb 04

Patanè, L;Cadamuro, M;Massazza, G;Pirola, S;Stagnati, V;Comerio, C;Carnelli, M;Arosio, M;Callegaro, AP;Tebaldi, P;Rigoli, E;Gianatti, A;Morotti, D;
PMID: 35131495 | DOI: 10.1016/j.ajogmf.2022.100589

Data on the vertical transmission rate of COVID-19 in pregnancy are limited, while data reporting mother-fetal transmission in the second trimester of pregnancy are controversial. We described a case of second trimester twin stillbirth in a woman positive for SARS-CoV-2 in which, despite the absence of respiratory syndrome, placental and fetal markers of infection were detected. The patient developed a clinical chorioamnionitis and spontaneously delivered two stillborn infants. Placental histology and immunohistochemistry demonstrated SARS-CoV-2 infection mostly within the syncytiotrophoblast and the fetal autopsy showed development of interstitial pneumonia. Our findings demonstrate that, in utero vertical transmission is possible, also in asymptomatic SARS-CoV-2 pregnant women and that infection can lead to severe morbidity in the second trimester of pregnancy.

Multisystem screening reveals SARS-CoV-2 in neurons of the myenteric plexus and in megakaryocytes

The Journal of pathology

2022 Feb 02

Gray-Rodriguez, S;Jensen, MP;Otero-Jimenez, M;Hanley, B;Swann, OC;Ward, PA;Salguero, FJ;Querido, N;Farkas, I;Velentza-Almpani, E;Weir, J;Barclay, WS;Carroll, MW;Jaunmuktane, Z;Brandner, S;Pohl, U;Allinson, K;Thom, M;Troakes, C;Al-Sarraj, S;Sastre, M;Gveric, D;Gentleman, S;Roufosse, C;Osborn, M;Alegre-Abarrategui, J;
PMID: 35107828 | DOI: 10.1002/path.5878

SARS-CoV-2, the causative agent of COVID-19, typically manifests as a respiratory illness although extrapulmonary involvement, such as in the gastrointestinal tract and nervous system, as well as frequent thrombotic events, are increasingly recognised. How this maps onto SARS-CoV-2 organ tropism at the histological level, however, remains unclear. Here, we perform a comprehensive validation of a monoclonal antibody against the SARS-CoV-2 nucleocapsid protein (NP) followed by systematic multisystem organ immunohistochemistry analysis of the viral cellular tropism in tissue from 36 patients, 16 post-mortem cases and 16 biopsies with polymerase chain reaction (PCR)-confirmed SARS-CoV-2 status from the peaks of the pandemic in 2020 and four pre-COVID post-mortem controls. SARS-CoV-2 anti-NP staining in the post-mortem cases revealed broad multiorgan involvement of the respiratory, digestive, haematopoietic, genitourinary and nervous systems, with a typical pattern of staining characterised by punctate paranuclear and apical cytoplasmic labelling. The average time from symptom onset to time of death was shorter in positively versus negatively stained post-mortem cases (mean = 10.3 days versus mean = 20.3 days, p = 0.0416, with no cases showing definitive staining if the interval exceeded 15 days). One striking finding was the widespread presence of SARS-CoV-2 NP in neurons of the myenteric plexus, a site of high ACE-2 expression, the entry receptor for SARS-CoV-2, and one of the earliest affected cells in Parkinson's disease. In the bone marrow, we observed viral SARS-CoV-2 NP within megakaryocytes, key cells in platelet production and thrombus formation. In 15 tracheal biopsies performed in patients requiring ventilation, there was a near complete concordance between immunohistochemistry and PCR swab results. Going forward, our findings have relevance to correlating clinical symptoms to the organ tropism of SARS-CoV-2 in contemporary cases as well as providing insights into potential long-term complications of COVID-19. This article is protected by

Long-term stability and protection efficacy of the RBD-targeting COVID-19 mRNA vaccine in nonhuman primates

Signal transduction and targeted therapy

2021 Dec 24

Zhao, H;Wang, TC;Li, XF;Zhang, NN;Li, L;Zhou, C;Deng, YQ;Cao, TS;Yang, G;Li, RT;Huang, YJ;Li, YG;Zhang, YM;Li, FX;Zhou, YR;Jiang, YH;Lu, XS;Sun, SH;Cheng, ML;Gu, KP;Zhang, M;Ma, QQ;Yang, X;Ying, B;Gao, YW;Qin, CF;
PMID: 34952914 | DOI: 10.1038/s41392-021-00861-4

Messenger RNA (mRNA) vaccine technology has shown its power in preventing the ongoing COVID-19 pandemic. Two mRNA vaccines targeting the full-length S protein of SARS-CoV-2 have been authorized for emergency use. Recently, we have developed a lipid nanoparticle-encapsulated mRNA (mRNA-LNP) encoding the receptor-binding domain (RBD) of SARS-CoV-2 (termed ARCoV), which confers complete protection in mouse model. Herein, we further characterized the protection efficacy of ARCoV in nonhuman primates and the long-term stability under normal refrigerator temperature. Intramuscular immunization of two doses of ARCoV elicited robust neutralizing antibodies as well as cellular response against SARS-CoV-2 in cynomolgus macaques. More importantly, ARCoV vaccination in macaques significantly protected animals from acute lung lesions caused by SARS-CoV-2, and viral replication in lungs and secretion in nasal swabs were completely cleared in all animals immunized with low or high doses of ARCoV. No evidence of antibody-dependent enhancement of infection was observed throughout the study. Finally, extensive stability assays showed that ARCoV can be stored at 2-8 °C for at least 6 months without decrease of immunogenicity. All these promising results strongly support the ongoing clinical trial.

AUTOPSY STUDY OF TESTICLES IN COVID-19: UPREGULATION OF IMMUNE-RELATED GENES AND DOWNREGULATION OF TESTIS-SPECIFIC GENES

The Journal of clinical endocrinology and metabolism

2022 Oct 19

Basolo, A;Poma, AM;Macerola, E;Bonuccelli, D;Proietti, A;Salvetti, A;Vignali, P;Torregrossa, L;Evangelisti, L;Sparavelli, R;Giannini, R;Ugolini, C;Basolo, F;Santini, F;Toniolo, A;
PMID: 36260523 | DOI: 10.1210/clinem/dgac608

Infection by SARS-CoV-2 may be associated with testicular dysfunction that could affect male fertility.Testicles of fatal COVID-19 cases were investigated to detect virus in tissue and to evaluate histopathological and transcriptomic changes.Three groups were compared: a. uninfected controls (subjects dying of trauma or sudden cardiac death; n = 10); b. subjects dying of COVID-19 (virus-negative in testes; n = 15); c. subjects dying of COVID-19 (virus-positive in testes; n = 9). SARS-CoV-2 genome and nucleocapsid antigen were probed using RT-PCR, in situ hybridization, immunohistochemistry (IHC). Infiltrating leukocytes were typed by IHC. mRNA transcripts of immune-related and testis-specific genes were quantified using the nCounter method.SARS-CoV-2 was detected in testis tissue of 9/24 (37%) COVID-19 cases accompanied by scattered T-cell and macrophage infiltrates. Size of testicles and counts of spermatogenic cells were not significantly different among groups. Analysis of mRNA transcripts showed that in virus-positive testes immune processes were activated (interferon-alpha and -gamma pathways). By contrast, transcription of 12 testis-specific genes was downregulated, independently of virus positivity in tissue. By IHC, expression of the luteinizing hormone/choriogonadotropin receptor was enhanced in virus-positive compared to virus-negative testicles, while expression of receptors for androgens and the follicle-stimulating hormone were not significantly different among groups.In lethal COVID-19 cases, infection of testicular cells is not uncommon. Viral infection associates with activation of interferon pathways and downregulation of testis-specific genes involved in spermatogenesis. Due to the exceedingly high numbers of infected people in the pandemic, the impact of virus on fertility should be further investigated.

Is thyroid gland a target of SARS-CoV-2 infection? Results of the analysis of necropsy thyroid specimens from COVID-19 patients

Endocrine Abstracts

2021 May 15

Macedo, S;Pestana, A;Liliana, R;Neves, C;Susana, G;Guimarães, A;Dolhnikoff, M;Saldiva, P;Carneiro, F;Sobrinho-Simões, M;Soares, P;
| DOI: 10.1530/endoabs.73.oc14.3

In the 2002 outbreak of severe acute respiratory syndrome (SARS) a number of patients presented abnormalities in the thyroid functioning, neuroendocrine and calcium homeostasis. It was detected in autopsies from SARS Coronavirus (SARS-CoV) patients that the thyroid gland was significantly affected by the disease, with extensive injury and death of follicular and parafollicular cells. In the present SARS-CoV-2 pandemic some studies start to report acute thyroiditis and alterations in the levels of thyroid hormones [(triiodothyronine (T3), thyroxine (T4), thyroid stimulating hormone (TSH)]. Thyroid cells present high levels of mRNA expression of angiotensin-converting enzyme 2 (ACE2), the host receptor for SARS-CoV-2. It remains poorly studied the thyroid expression of proteins that predispose to SARS-CoV-2 infection and if thyroid cells can be a direct or indirect target of SARS-CoV-2 infection.

Subcellular Detection of SARS-CoV-2 RNA in Human Tissue Reveals Distinct Localization in Alveolar Type 2 Pneumocytes and Alveolar Macrophages

mBio

2022 Feb 08

Acheampong, KK;Schaff, DL;Emert, BL;Lake, J;Reffsin, S;Shea, EK;Comar, CE;Litzky, LA;Khurram, NA;Linn, RL;Feldman, M;Weiss, SR;Montone, KT;Cherry, S;Shaffer, SM;
PMID: 35130722 | DOI: 10.1128/mbio.03751-21

The widespread coronavirus disease 2019 (COVID-19) is caused by infection with the novel coronavirus SARS-CoV-2. Currently, we have limited understanding of which cells become infected with SARS-CoV-2 in human tissues and where viral RNA localizes on the subcellular level. Here, we present a platform for preparing autopsy tissue for visualizing SARS-CoV-2 RNA using RNA fluorescence in situ hybridization (FISH) with amplification by hybridization chain reaction. We developed probe sets that target different regions of SARS-CoV-2 (including ORF1a and N), as well as probe sets that specifically target SARS-CoV-2 subgenomic mRNAs. We validated these probe sets in cell culture and tissues (lung, lymph node, and placenta) from infected patients. Using this technology, we observe distinct subcellular localization patterns of the ORF1a and N regions. In human lung tissue, we performed multiplexed RNA FISH HCR for SARS-CoV-2 and cell-type-specific marker genes. We found viral RNA in cells containing the alveolar type 2 (AT2) cell marker gene (SFTPC) and the alveolar macrophage marker gene (MARCO) but did not identify viral RNA in cells containing the alveolar type 1 (AT1) cell marker gene (AGER). Moreover, we observed distinct subcellular localization patterns of viral RNA in AT2 cells and alveolar macrophages. In sum, we demonstrate the use of RNA FISH HCR for visualizing different RNA species from SARS-CoV-2 in cell lines and FFPE (formalin fixation and paraffin embedding) autopsy specimens. We anticipate that this platform could be broadly useful for studying SARS-CoV-2 pathology in tissues, as well as extended for other applications, including investigating the viral life cycle, viral diagnostics, and drug screening. IMPORTANCE Here, we developed an in situ RNA detection assay for RNA generated by the SARS-CoV-2 virus. We found viral RNA in lung, lymph node, and placenta samples from pathology specimens from COVID patients. Using high-magnification microscopy, we can visualize the subcellular distribution of these RNA in single cells.

Neonates and COVID-19: state of the art: Neonatal Sepsis series

Pediatric research

2021 Dec 28

Ryan, L;Plötz, FB;van den Hoogen, A;Latour, JM;Degtyareva, M;Keuning, M;Klingenberg, C;Reiss, IKM;Giannoni, E;Roehr, C;Gale, C;Molloy, EJ;
PMID: 34961785 | DOI: 10.1038/s41390-021-01875-y

The SARS-CoV-2 pandemic has had a significant impact worldwide, particularly in middle- and low-income countries. While this impact has been well-recognized in certain age groups, the effects, both direct and indirect, on the neonatal population remain largely unknown. There are placental changes associated, though the contributions to maternal and fetal illness have not been fully determined. The rate of premature delivery has increased and SARS-CoV-2 infection is proportionately higher in premature neonates, which appears to be related to premature delivery for maternal reasons rather than an increase in spontaneous preterm labor. There is much room for expansion, including long-term data on outcomes for affected babies. Though uncommon, there has been evidence of adverse events in neonates, including Multisystem Inflammatory Syndrome in Children, associated with COVID-19 (MIS-C). There are recommendations for reduction of viral transmission to neonates, though more research is required to determine the role of passive immunization of the fetus via maternal vaccination. There is now considerable evidence suggesting that the severe visitation restrictions implemented early in the pandemic have negatively impacted the care of the neonate and the experiences of both parents and healthcare professionals alike. Ongoing collaboration is required to determine the full impact, and guidelines for future management. IMPACT: Comprehensive review of current available evidence related to impact of the COVID-19 pandemic on neonates, effects on their health, impact on their quality of care and indirect influences on their clinical course, including comparisons with other age groups. Reference to current evidence for maternal experience of infection and how it impacts the fetus and then neonate. Outline of the need for ongoing research, including specific areas in which there are significant gaps in knowledge.

Acute Mesenteric Ischemia in Patients with COVID-19: Review of the literature

Journal of the National Medical Association

2021 Dec 29

Chen, C;Li, YW;Shi, PF;Qian, SX;
PMID: 34973847 | DOI: 10.1016/j.jnma.2021.12.003

The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has resulted in a global health emergency. In addition to common respiratory symptoms, some patients with COVID-19 infections may experience a range of extra-pulmonary manifestations, such as digestive system involvement. Patients with COVID-19 have been reported to suffer from acute mesenteric ischemia (AMI) that is associated with disease-related severity and mortality. However, in the context of COVID-19, the exact cause of AMI has yet to be clearly defined. This review provides a comprehensive overview of the available data and elucidates the possible underlying mechanisms linking COVID-19 to AMI, in addition to highlighting therapeutic approaches for clinicians. Finally, given the severe global impact of COVID-19, we emphasize the importance of coordinated vaccination programs.

Tissue-Based SARS-Cov-2 Detection in Fatal COVID-19 Infections: Sustained Direct Viral-Induced Damage is Not Necessary to Drive Disease Progression

Human pathology

2021 May 04

El Jamal, SM;Pujadas, E;Ramos, I;Bryce, C;Grimes, ZM;Amanat, F;Tsankova, NM;Mussa, Z;Olson, S;Salem, F;Miorin, L;Aydillo, T;Schotsaert, M;Albrecht, RA;Liu, WC;Marjanovic, N;Francoeur, N;Sebra, R;Sealfon, SC;García-Sastre, A;Fowkes, M;Cordon-Cardo, C;Westra, WH;
PMID: 33961839 | DOI: 10.1016/j.humpath.2021.04.012

Coronavirus disease 2019 (COVID-19) is an ongoing pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Although viral infection is known to trigger inflammatory processes contributing to tissue injury and organ failure, it is unclear whether direct viral damage is needed to sustain cellular injury. An understanding of pathogenic mechanisms has been handicapped by the absence of optimized methods to visualize the presence and distribution of SARS-CoV-2 in damaged tissues. We first developed a positive control cell line (Vero E6) to validate SARS-CoV-2 detection assays. We then evaluated multiple organs (lungs, kidneys, heart, liver, brain, intestines, lymph nodes and spleen) from fourteen COVID-19 autopsy cases using immunohistochemistry (IHC) for the spike and the nucleoprotein proteins, and RNA in-situ hybridization (RNA ISH) for the spike protein mRNA. Tissue detection assays were compared with quantitative PCR (qPCR)-based detection. SARS-CoV-2 was histologically detected in the Vero E6 positive cell line control, 1 of 14 (7%) lungs, and none (0%) of the other 59 organs. There was perfect concordance between the IHC and RNA ISH results. qPCR confirmed high viral load in the SARS-CoV-2 ISH-positive lung tissue, and absent or low viral load in all ISH-negative tissues. In patients who die of COVID-19-related organ failure, SARS-CoV-2 is largely not detectable using tissue-based assays. Even in lungs showing widespread injury, SARS-CoV-2 viral RNA or proteins were detected in only a small minority of cases. This observation supports the concept that viral infection is primarily a trigger for multiple organ pathogenic pro-inflammatory responses. Direct viral tissue damage is a transient phenomenon that is generally not sustained throughout disease progression.

Immune-guided therapy of COVID-19

Cancer immunology research

2022 Jan 24

Ferraccioli, G;Gremese, E;Goletti, D;Petrone, L;Cantini, F;Ugel, S;Canè, S;Bronte, V;
PMID: 35074758 | DOI: 10.1158/2326-6066.CIR-21-0675

Vaccination has been a game changer in our efforts to address the coronavirus disease 2019 (COVID-19) pandemic. However, the disease might still represent a clinical crisis for several more years, in part because of the inevitable emergence of variants capable of evading the pre-existing immunity. Drugs affecting viral spread will help curtail transmission, but therapeutics are needed to treat the more severe cases requiring hospitalization. A deep analysis of the evolving immune landscape of COVID-19 suggests that understanding the molecular bases of the distinct clinical stages is paramount if we are to limit the burden of inflammation, which can lead to death in frail individuals, according to age, sex and comorbidities. Different phases can be defined using immune biomarkers and need specific therapeutic approaches, tailored to the underlying immune contexture.

LB981 Pandemic associated chilblain-like lesions result from an inducible type 1 interferon response to SARS-CoV-2

Journal of Investigative Dermatology

2022 Aug 01

Arkin, L;Costa da Silva, A;Mays, J;
| DOI: 10.1016/j.jid.2022.05.1004

Chilblain-like lesions (CLL), known in the lay press as “COVID toes,” increased significantly during the COVID-19 pandemic. The phenotypic similarity of chilblains in the monogenic type 1 interferonopathies, coupled with the consistent clinical phenotype across multiple countries and temporospatial association with COVID-19 spread, suggest a SARS-CoV-2 triggered immune phenomenon. Yet direct evidence of this relationship has been limited due to low rates of SARS-CoV-2 positivity utilizing conventional testing. We prospectively enrolled a cohort of 79 patients with CLL across 4 waves of the SARS-CoV-2 pandemic in Wisconsin collecting serial blood samples and lesional skin biopsies. Immunophenotyping including the type 1 interferon (IFN-1) signature was investigated utilizing multiplex immunohistochemistry in affected tissue. Proteomics and RNA sequencing were performed on the peripheral blood at serial time points. RNAscope for S gene and depositional immunohistochemistry for evidence of SARS-CoV-2 were performed on tissue. Antibody responses and T-cell specific responses to SARS-CoV-2 were performed and an animal model (golden hamster) provided mechanistic evidence of dissemination of viral RNA to acral sites with local IFN-1 activation. Our results support an inducible local and peripheral IFN-1 signature, which abrogates within weeks, with evidence of viral SARS-CoV-2 RNA as the trigger.

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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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