Exosomes decorated with a recombinant SARS-CoV-2 receptor-binding domain as an inhalable COVID-19 vaccine

Nature biomedical engineering

2022 Jul 01

Wang, Z;Popowski, KD;Zhu, D;de Juan Abad, BL;Wang, X;Liu, M;Lutz, H;De Naeyer, N;DeMarco, CT;Denny, TN;Dinh, PC;Li, Z;Cheng, K;
PMID: 35788687 | DOI: 10.1038/s41551-022-00902-5

The first two mRNA vaccines against infection by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that were approved by regulators require a cold chain and were designed to elicit systemic immunity via intramuscular injection. Here we report the design and preclinical testing of an inhalable virus-like-particle as a COVID-19 vaccine that, after lyophilisation, is stable at room temperature for over three months. The vaccine consists of a recombinant SARS-CoV-2 receptor-binding domain (RBD) conjugated to lung-derived exosomes which, with respect to liposomes, enhance the retention of the RBD in both the mucus-lined respiratory airway and in lung parenchyma. In mice, the vaccine elicited RBD-specific IgG antibodies, mucosal IgA responses and CD4+ and CD8+ T cells with a Th1-like cytokine expression profile in the animals' lungs, and cleared them of SARS-CoV-2 pseudovirus after a challenge. In hamsters, two doses of the vaccine attenuated severe pneumonia and reduced inflammatory infiltrates after a challenge with live SARS-CoV-2. Inhalable and room-temperature-stable virus-like particles may become promising vaccine candidates.

Nonhuman primate models for evaluation of SARS-CoV-2 vaccines

Expert review of vaccines

2022 Jun 02

Neil, JA;Griffith, M;Godfrey, DI;Purcell, DFJ;Deliyannis, G;Jackson, D;Rockman, S;Subbarao, K;Nolan, T;
PMID: 35652289 | DOI: 10.1080/14760584.2022.2071264

Evaluation of immunogenicity and efficacy in animal models provide critical data in vaccine development. Nonhuman primates (NHPs) have been used extensively in the evaluation of SARS-CoV-2 vaccines.A critical synthesis of SARS-CoV-2 vaccine development with a focus on challenge studies in NHPs is provided. The benefits and drawbacks of the NHP models are discussed. The citations were selected by the authors based on PubMed searches of the literature, summaries from national public health bodies, and press-release information provided by vaccine developers.We identify several aspects of NHP models that limit their usefulness for vaccine-challenge studies and numerous variables that constrain comparisons across vaccine platforms. We propose that studies conducted in NHPs for vaccine development should use a standardized protocol and, where possible, be substituted with smaller animal models. This will ensure continued rapid progression of vaccines to clinical trials without compromising assessments of safety or efficacy.

Cell tropism and viral clearance during SARS-CoV-2 lung infection

Pathology - Research and Practice

2022 Jun 01

Schwab, C;Domke, L;Rose, F;Hausser, I;Schirmacher, P;Longerich, T;
| DOI: 10.1016/j.prp.2022.154000

Pulmonary capillary microthrombosis has been proposed as a major pathogenetic factor driving severe COVID-19. Autopsy studies reported endothelialitis but it is under debate if it is caused by SARS-CoV-2 infection of endothelial cells. In this study, RNA in situ hybridization was used to detect viral RNA and to identify the infected cell types in lung tissue of 40 patients with fatal COVID-19. SARS-CoV-2 Spike protein-coding RNA showed a steadily decreasing signal abundance over a period of three weeks. Besides the original virus strain the variants of concern Alpha (B.1.1.7), Delta (B.1.617.2), and Omicron (B.1.1.529) could also be detected by the assay. Viral RNA was mainly detected in alveolar macrophages and pulmonary epithelial cells, while only single virus-positive endothelial cells were observed even in cases with high viral load suggesting that viral infection of endothelial cells is not a key factor for the development of pulmonary capillary microthrombosis.

SARS-CoV-2 treatment effects induced by ACE2-expressing microparticles are explained by the oxidized cholesterol-increased endosomal pH of alveolar macrophages

Cellular & molecular immunology

2022 Feb 01

Wang, Z;Lv, J;Yu, P;Qu, Y;Zhou, Y;Zhou, L;Zhu, Q;Li, S;Song, J;Deng, W;Gao, R;Liu, Y;Liu, J;Tong, WM;Qin, C;Huang, B;
PMID: 34983944 | DOI: 10.1038/s41423-021-00813-6

Exploring the cross-talk between the immune system and advanced biomaterials to treat SARS-CoV-2 infection is a promising strategy. Here, we show that ACE2-overexpressing A549 cell-derived microparticles (AO-MPs) are a potential therapeutic agent against SARS-CoV-2 infection. Intranasally administered AO-MPs dexterously navigate the anatomical and biological features of the lungs to enter the alveoli and are taken up by alveolar macrophages (AMs). Then, AO-MPs increase the endosomal pH but decrease the lysosomal pH in AMs, thus escorting bound SARS-CoV-2 from phago-endosomes to lysosomes for degradation. This pH regulation is attributable to oxidized cholesterol, which is enriched in AO-MPs and translocated to endosomal membranes, thus interfering with proton pumps and impairing endosomal acidification. In addition to promoting viral degradation, AO-MPs also inhibit the proinflammatory phenotype of AMs, leading to increased treatment efficacy in a SARS-CoV-2-infected mouse model without side effects. These findings highlight the potential use of AO-MPs to treat SARS-CoV-2-infected patients and showcase the feasibility of MP therapies for combatting emerging respiratory viruses in the future.

Neuropsychiatric manifestations of COVID-19, potential neurotropic mechanisms, and therapeutic interventions

Translational psychiatry

2021 Sep 30

Han, Y;Yuan, K;Wang, Z;Liu, WJ;Lu, ZA;Liu, L;Shi, L;Yan, W;Yuan, JL;Li, JL;Shi, J;Liu, ZC;Wang, GH;Kosten, T;Bao, YP;Lu, L;
PMID: 34593760 | DOI: 10.1038/s41398-021-01629-8

The coronavirus disease 2019 (COVID-19) pandemic has caused large-scale economic and social losses and worldwide deaths. Although most COVID-19 patients have initially complained of respiratory insufficiency, the presence of neuropsychiatric manifestations is also reported frequently, ranging from headache, hyposmia/anosmia, and neuromuscular dysfunction to stroke, seizure, encephalopathy, altered mental status, and psychiatric disorders, both in the acute phase and in the long term. These neuropsychiatric complications have emerged as a potential indicator of worsened clinical outcomes and poor prognosis, thus contributing to mortality in COVID-19 patients. Their etiology remains largely unclear and probably involves multiple neuroinvasive pathways. Here, we summarize recent animal and human studies for neurotrophic properties of severe acute respiratory syndrome coronavirus (SARS-CoV-2) and elucidate potential neuropathogenic mechanisms involved in the viral invasion of the central nervous system as a cause for brain damage and neurological impairments. We then discuss the potential therapeutic strategy for intervening and preventing neuropsychiatric complications associated with SARS-CoV-2 infection. Time-series monitoring of clinical-neurochemical-radiological progress of neuropsychiatric and neuroimmune complications need implementation in individuals exposed to SARS-CoV-2. The development of a screening, intervention, and therapeutic framework to prevent and reduce neuropsychiatric sequela is urgently needed and crucial for the short- and long-term recovery of COVID-19 patients.

LB740 SARS-CoV-2-associated ‘covid toes:’ multiplex immunofluorescent characterization of pathophysiology

Journal of Investigative Dermatology

2021 Sep 01

Moon, J;Costa da Silva, A;Tran, J;Kim, C;Sharma, R;Hinshaw, M;Shields, B;Brooks, E;Cowen, E;Singh, A;Drolet, B;Mays, J;Arkin, L;
| DOI: 10.1016/j.jid.2021.07.093

Coincident with the start of the COVID-19 pandemic, dermatologists worldwide have reported an uncharacteristic increase in pernio or chilblains (aka ‘COVID toes’). However, the lack of systemic illness, low PCR positivity and lack of consistent seroconversion have led some authors to postulate an epiphenomenon. SARS-CoV-2 spike protein has been identified in a limited number of skin biopsies in few publications, yet there remain conflicting reports regarding other SARS-CoV-2 associated proteins, the presence or absence of viral RNA, and a unifying pathophysiology. In cooperation with the COVID Human Genome Effort, our “COVID toes” biobank was established to identify both the genetic and immunologic basis and provide clinically relevant insights into targeted therapeutics. As of March 2021, we have enrolled 96 patients, creating a prospective biorepository with clinical data, saliva, serial blood collection, and skin biopsies. Here we aim to comprehensively investigate the conflicting findings, detail the inflammatory response, and identify the source of interferon signaling with multiplex immunofluorescence (IFA) and the RNAscope fluorescent assay to detect viral mRNA. Median patient age was 17 (range 2 e 72) and 44/96 (46%) were male. Preliminary IFA results demonstrate detection of SARS-CoV-2 components, robust MxA detection and plasmacytoid dendritic cell (pDC) colocalization, identifying PDCs as the likely primary source of IFN-I production and implicates an excessive localized IFN-I response in affected patients.

SARS-CoV-2 Omicron virus causes attenuated disease in mice and hamsters

Nature

2022 Jan 21

Halfmann, PJ;Iida, S;Iwatsuki-Horimoto, K;Maemura, T;Kiso, M;Scheaffer, SM;Darling, TL;Joshi, A;Loeber, S;Singh, G;Foster, SL;Ying, B;Case, JB;Chong, Z;Whitener, B;Moliva, J;Floyd, K;Ujie, M;Nakajima, N;Ito, M;Wright, R;Uraki, R;Warang, P;Gagne, M;Li, R;Sakai-Tagawa, Y;Liu, Y;Larson, D;Osorio, JE;Hernandez-Ortiz, JP;Henry, AR;Ciouderis, K;Florek, KR;Patel, M;Odle, A;Wong, LR;Bateman, AC;Wang, Z;Edara, VV;Chong, Z;Franks, J;Jeevan, T;Fabrizio, T;DeBeauchamp, J;Kercher, L;Seiler, P;Gonzalez-Reiche, AS;Sordillo, EM;Chang, LA;van Bakel, H;Simon, V;Consortium Mount Sinai Pathogen Surveillance (PSP) study group, ;Douek, DC;Sullivan, NJ;Thackray, LB;Ueki, H;Yamayoshi, S;Imai, M;Perlman, S;Webby, RJ;Seder, RA;Suthar, MS;García-Sastre, A;Schotsaert, M;Suzuki, T;Boon, ACM;Diamond, MS;Kawaoka, Y;
PMID: 35062015 | DOI: 10.1038/s41586-022-04441-6

The recent emergence of B.1.1.529, the Omicron variant1,2 has raised concerns for escape from protection by vaccines and therapeutic antibodies. A key test for potential countermeasures against B.1.1.529 is their activity in pre-clinical rodent models of respiratory tract disease. Here, using the collaborative network of the SARS-CoV-2 Assessment of Viral Evolution (SAVE) program of the National Institute of Allergy and Infectious Diseases (NIAID), we evaluated the ability of multiple B.1.1.529 Omicron isolates to cause infection and disease in immunocompetent and human ACE2 (hACE2) expressing mice and hamsters. Despite modeling data suggesting that B.1.1.529 spike can bind more avidly to murine ACE23,4, we observed less infection in 129, C57BL/6, BALB/c, and K18-hACE2 transgenic mice as compared with previous SARS-CoV-2 variants, with limited weight loss and lower viral burden in the upper and lower respiratory tracts. In wild-type and hACE2 transgenic hamsters, lung infection, clinical disease, and pathology with B.1.1.529 also were milder compared to historical isolates or other SARS-CoV-2 variants of concern. Overall, experiments from the SAVE/NIAID network with several B.1.1.529 isolates demonstrate attenuated lung disease in rodents, which parallels preliminary human clinical data.

Bilateral Chilblain-like Lesions of the Toes Characterized by Microvascular Remodeling in Adolescents During the COVID-19 Pandemic

JAMA network open

2021 Jun 01

Discepolo, V;Catzola, A;Pierri, L;Mascolo, M;Della Casa, F;Vastarella, M;Smith, G;Travaglino, A;Punziano, A;Nappa, P;Staibano, S;Bruzzese, E;Fabbrocini, G;Guarino, A;Alessio, M;
PMID: 34110396 | DOI: 10.1001/jamanetworkopen.2021.11369

Chilblain-like lesions have been one of the most frequently described cutaneous manifestations during the COVID-19 pandemic. Their etiopathogenesis, including the role of SARS-CoV-2, remains elusive.To examine the association of chilblain-like lesions with SARS-CoV-2 infection.This prospective case series enrolled 17 adolescents who presented with chilblain-like lesions from April 1 to June 30, 2020, at a tertiary referral academic hospital in Italy.Macroscopic (clinical and dermoscopic) and microscopic (histopathologic) analysis contributed to a thorough understanding of the lesions. Nasopharyngeal swab, serologic testing, and in situ hybridization of the skin biopsy specimens were performed to test for SARS-CoV-2 infection. Laboratory tests explored signs of systemic inflammation or thrombophilia. Structural changes in peripheral microcirculation were investigated by capillaroscopy.Of the 17 adolescents (9 [52.9%] male; median [interquartile range] age, 13.2 [12.5-14.3] years) enrolled during the first wave of the COVID-19 pandemic, 16 (94.1%) had bilaterally localized distal erythematous or cyanotic lesions. A triad of red dots (16 [100%]), white rosettes (11 [68.8%]), and white streaks (10 [62.5%]) characterized the dermoscopic picture. Histologic analysis revealed a remodeling of the dermal blood vessels with a lobular arrangement, wall thickening, and a mild perivascular lymphocytic infiltrate. SARS-CoV-2 infection was excluded by molecular and serologic testing. In situ hybridization did not highlight the viral genome in the lesions.This study delineated the clinical, histologic, and laboratory features of chilblain-like lesions that emerged during the COVID-19 pandemic, and its findings do not support their association with SARS-CoV-2 infection. The lesions occurred in otherwise healthy adolescents, had a long but benign course to self-resolution, and were characterized by a microvascular remodeling with perivascular lymphocytic infiltrate but no other signs of vasculitis. These results suggest that chilblain-like lesions do not imply a concomitant SARS-CoV-2 infection. Ongoing studies will help clarify the etiopathogenic mechanisms.

Hamsters Expressing Human Angiotensin-Converting Enzyme 2 Develop Severe Disease following Exposure to SARS-CoV-2

mBio

2022 Jan 25

Golden, JW;Li, R;Cline, CR;Zeng, X;Mucker, EM;Fuentes-Lao, AJ;Spik, KW;Williams, JA;Twenhafel, N;Davis, N;Moore, JL;Stevens, S;Blue, E;Garrison, AR;Larson, DD;Stewart, R;Kunzler, M;Liu, Y;Wang, Z;Hooper, JW;
PMID: 35073750 | DOI: 10.1128/mbio.02906-21

The rapid emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has created a global health emergency. While most human disease is mild to moderate, some infections lead to a severe disease characterized by acute respiratory distress, hypoxia, anosmia, ageusia, and, in some instances, neurological involvement. Small-animal models reproducing severe disease, including neurological sequela, are needed to characterize the pathophysiological mechanism(s) of disease and to identify medical countermeasures. Transgenic mice expressing the human angiotensin-converting enzyme 2 (hACE2) viral receptor under the control of the K18 promoter develop severe and lethal respiratory disease subsequent to SARS-CoV-2 intranasal challenge when high viral doses are used. Here, we report on SARS-CoV-2 infection of hamsters engineered to express the hACE2 receptor under the control of the K18 promoter. K18-hACE2 hamsters infected with a relatively low dose of 100 or 1,000 PFU of SARS-CoV-2 developed a severe and lethal disease, with most animals succumbing by day 5 postinfection. Hamsters developed severe lesions and inflammation within the upper and lower respiratory system, including infection of the nasal cavities causing marked destruction of the olfactory epithelium as well as severe bronchopneumonia that extended deep into the alveoli. Additionally, SARS-CoV-2 infection spread to the central nervous system (CNS), including the brain stem and spinal cord. Wild-type (WT) hamsters naturally support SARS-CoV-2 infection, with the primary lesions present in the respiratory tract and nasal cavity. Overall, infection in the K18-hACE2 hamsters is more extensive than that in WT hamsters, with more CNS involvement and a lethal outcome. These findings demonstrate the K18-hACE2 hamster model will be valuable for studying SARS-CoV-2. IMPORTANCE The rapid emergence of SARS-CoV-2 has created a global health emergency. While most human SARS-CoV-2 disease is mild, some people develop severe, life-threatening disease. Small-animal models mimicking the severe aspects of human disease are needed to more clearly understand the pathophysiological processes driving this progression. Here, we studied SARS-CoV-2 infection in hamsters engineered to express the human angiotensin-converting enzyme 2 viral receptor under the control of the K18 promoter. SARS-CoV-2 produces a severe and lethal infection in transgenic hamsters that mirrors the most severe aspects of COVID-19 in humans, including respiratory and neurological injury. In contrast to other animal systems, hamsters manifest disease with levels of input virus more consistent with natural human infection. This system will be useful for the study of SARS-CoV-2 disease and the development of drugs targeting this virus.

Postmortem Minimally Invasive Autopsy in Critically Ill Covid-19 Patients at the Bedside: A Proof-of-Concept Study at the Icu

SSRN

2023 Jun 02

Lahmer, T;Weirich, G;Porubsky, S;Rasch, S;Kammerstetter, F;Schustetter, C;Schüffler, P;Erber, J;Dibos, M;delbridge, c;kuhn, p;Jeske, S;steinhardt, m;Chaker, A;Heim, M;Heemann, U;Schmid, R;weichert, W;Stock, K;Slotta, J;
| DOI: 10.2139/ssrn.4464818

Methods: In this proof-of-concept study, we performed bedside ultrasound-guided minimally invasive autopsies (US-MIA) of patients that had died from critical COVID-19 in the intensive care unit (ICU) using a structured protocol to obtain almost autolytic-free tissue. Biopsies were assessed for quality (vitality and length) and for diagnosis. The efficiency of the procedure was monitored in five cases by recording the time of each step and safety issues by swabbing personal protective equipment and devices for viral contamination.

Diagnostic role of technitium-99m bone scan in severe COVID-19-associated myositis

Radiology case reports

2021 May 14

Hookey, G;Ahmad, Q;McCune, T;Kowalewska, J;Amaker, B;Inayat, N;
PMID: 34007372 | DOI: 10.1016/j.radcr.2021.05.019

Coronavirus disease 2019 (COVID-19), initially appreciated as a respiratory illness, is now known to affect many organs in the human body. Significant data has become available on muscle involvement, with creatinine kinase elevations present in a significant percentage of patients. For those with suspected COVID-19-associated myositis, the imaging modality of choice has been gadolinium-enhanced magnetic resonance imaging; however, the use of technitium-99m bone scan has not been previously reported. Here, we report two cases of COVID-19 patients with severe elevation in creatinine kinase who underwent technitium-99m bone scan. The resulting images showed diffuse symmetrical muscle involvement. Both patients developed acute renal injury due to rhabdomyolysis. To our knowledge, this is the first report of bone scan as a diagnostic imaging modality for COVID-19-associated myositis.

Lung transplantation for coronavirus disease 2019 associated severe acute respiratory distress syndrome

Current opinion in anaesthesiology

2022 Oct 28

Garza-Castillon, R;Bharat, A;
PMID: 36302203 | DOI: 10.1097/ACO.0000000000001203

The purpose of this review is to analyze the most recent and relevant literature involving lung transplantation for coronavirus disease 2019 (COVID-19) associated acute respiratory distress syndrome (ARDS), the pathological mechanisms of lung injury, selection criteria and outcomes.Pathological analysis of lungs after COVID-19 ARDS has shown architectural distortion similar to that observed in explanted lungs from patients undergoing lung transplantation for end-stage lung diseases such as emphysema. Short-term outcomes after lung transplantation for COVID-19 associated respiratory failure are comparable to those performed for other indications.Lung transplantation after COVID-19 ARDS is a potentially life-saving procedure for appropriately selected patients with no evidence of lung function recovery despite maximal treatment. Lung transplantation should be ideally performed in high-volume centers with expertise.

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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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