BSL2-compliant lethal mouse model of SARS-CoV-2 and variants of concern to evaluate therapeutics targeting the Spike protein

Since first reported in 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is rapidly acquiring mutations, particularly in the spike protein, that can modulate pathogenicity, transmission and antibody evasion leading to successive waves of COVID19 infections despite an unprecedented mass vaccination necessitating continuous adaptation of therapeutics. Small animal models can facilitate understanding host-pathogen interactions, target selection for therapeutic drugs, and vaccine development, but availability and cost of studies in BSL3 facilities hinder progress. To generate a BSL2-compatibl

BSL2-compliant lethal mouse model of SARS-CoV-2 and variants of concern to evaluate therapeutics targeting the Spike protein

Since first reported in 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is rapidly acquiring mutations, particularly in the spike protein, that can modulate pathogenicity, transmission and antibody evasion leading to successive waves of COVID19 infections despite an unprecedented mass vaccination necessitating continuous adaptation of therapeutics. Small animal models can facilitate understanding host-pathogen interactions, target selection for therapeutic drugs, and vaccine development, but availability and cost of studies in BSL3 facilities hinder progress. To generate a BSL2-compatible in vivo system that specifically recapitulates spike protein mediated disease we used replication competent, GFP tagged, recombinant Vesicular Stomatitis Virus where the VSV glycoprotein was replaced by the SARS-CoV-2 spike protein (rVSV-SARS2-S). We show that infection requires hACE2 and challenge of neonatal but not adult, K18-hACE2 transgenic mice (hACE2tg) leads to productive infection of the lungs and brains. Although disease progression was faster in SARS-CoV-2 infected mice, infection with both viruses resulted in neuronal infection and encephalitis with increased expression of Interferon-stimulated Irf7, Bst2, Ifi294, as well as CxCL10, CCL5, CLC2, and LILRB4, and both models were uniformly lethal. Further, prophylactic treatment targeting the Spike protein (Receptor Binding Domain) with antibodies resulted in similar levels of protection from lethal infection against rVSV-SARS2-S and SARS-CoV-2 viruses. Strikingly, challenge of neonatal hACE2tg mice with SARS-CoV-2 Variants of Concern (SARS-CoV-2-α, -β, ϒ, or Δ) or the corresponding rVSV-SARS2-S viruses (rVSV-SARS2-Spike-α, rVSV-SARS2-Spike-β, rVSV-SARS2-Spike-ϒ or rVSV-SARS2-Spike-Δ) resulted in increased lethality, suggesting that the Spike protein plays a key role in determining the virulence of each variant. Thus, we propose that rVSV-SARS2-S virus can be used to understand the effect of changes to SARS-CoV-2 spike protein on infection and to evaluate existing or experimental therapeutics targeting spike protein of current or future VOC of SARS-CoV-2 under BSL-2 conditions

Repeated Low Intensity Blast Exposure Is Associated with Damaged Endothelial Glycocalyx and Downstream Behavioral Deficits

Current clinical research into mild traumatic brain injury (mTBI) has focused on white matter changes as identified by advanced MRI based imaging techniques. However, perivascular tau accumulation in the brains of individuals diagnosed with mTBI suggests that the vasculature plays a key role in the pathology. This study used a rat model to examine whether the endothelial glycocalyx, a layer of the vasculature responsible for sensing luminal shear forces, is damaged by exposure to repeated low intensity blast, and whether this layer is associated with observed behavioral deficits. The blast exposure used consisted of 12, 40 kPa blast exposures conducted with a minimum of 24 h between blasts. We found that repeated blast exposure reduced glycocalyx length and density in various brain regions indicating damage. This blast exposure paradigm was associated with a mild performance decrement in the Morris water maze (MWM) which assesses learning and memory. Administration of hyaluronidase, an enzyme that binds to and degrades hyaluronan (a major structural component of the glycocalyx) prior to blast exposure reduced the observed behavioral deficits and induced a thickening of the glycocalyx layer. Taken together these findings demonstrate that the endothelial glycocalyx degradation following repeated blast is associated with behavioral decrements which can be prevented by treatment with hyaluronidase

Socioeconomic, Clinical, and Molecular Features of Breast Cancer Influence Overall Survival of Latin American Women

Molecular profile of breast cancer in Latin-American women was studied in five countries: Argentina, Brazil, Chile, Mexico, and Uruguay. Data about socioeconomic characteristics, risk factors, prognostic factors, and molecular subtypes were described, and the 60-month overall cumulative survival probabilities (OS) were estimated. From 2011 to 2013, 1,300 eligible Latin-American women 18 years or older, with a diagnosis of breast cancer in clinical stage II or III, and performance status ≦̸1 were invited to participate in a prospective cohort study. Face-to-face interviews were conducted, and clinical and outcome data, including death, were extracted from medical records. Unadjusted associations were evaluated by Chi-squared and Fisher’s exact tests and the OS by Kaplan–Meier method. Log-rank test was used to determine differences between cumulative probability curves. Multivariable adjustment was carried out by entering potential confounders in the Cox regression model. The OS at 60 months was 83.9%. Multivariable-adjusted death hazard differences were found for women living in Argentina (2.27), Chile (1.95), and Uruguay (2.42) compared with Mexican women, for older (≥60 years) (1.84) compared with younger (≤40 years) women, for basal-like subtype (5.8), luminal B (2.43), and HER2-enriched (2.52) compared with luminal A subtype, and for tumor clinical stages IIB (1.91), IIIA (3.54), and IIIB (3.94) compared with stage IIA women. OS was associated with country of residence, PAM50 intrinsic subtype, age, and tumor stage at diagnosis. While the latter is known to be influenced by access to care, including cancer screening, timely diagnosis and treatment, including access to more effective treatment protocols, it may also influence epigenetic changes that, potentially, impact molecular subtypes. Data derived from heretofore understudied populations with unique geographic ancestry and sociocultural experiences are critical to furthering our understanding of this complexity. Copyright © 2022 de Almeida, Cortés, Vilensky, Valenzuela, Cortes-Sanabria, de Souza, Barbeito, Abdelhay, Artagaveytia, Daneri-Navarro, Llera, Müller, Podhajcer, Velazquez, Alcoba, Alonso, Bravo, Camejo, Carraro, Castro, Cataldi, Cayota, Cerda, Colombo, Crocamo, Del Toro-Arreola, Delgadillo-Cristerna, Delgado, Breitenbach, Fernández, Fernández, Fernández, Franco-Topete, Gaete, Gómez, Gonzalez-Ramirez, Guerrero, Gutierrez-Rubio, Jalfin, Lopez-Vazquez, Loria, Míguez, Moran-Mendoza, Morgan-Villela, Mussetti, Nagai, Oceguera-Villanueva, Reis, Retamales, Rodriguez, Rosales, Salas-Gonzalez, Segovia, Sendoya, Silva-Garcia, Viña, Zagame, Jones, Szklo and United States-Latin American Cancer Research Network (US-LACRN).Fil: de Almeida, Liz Maria. Instituto Nacional de Cáncer; BrasilFil: Cortés, Sandra. Pontificia Universidad Católica de Chile; ChileFil: Vilensky, Marta. Instituto de Oncología Angel Roffo; ArgentinaFil: Valenzuela, Olivia. Universidad de Sonora; MéxicoFil: Cortes-Sanabria, Laura. Hospital de Especialidades, CMNO-IMSS; MéxicoFil: de Souza, Mirian. Instituto Nacional de Cáncer; BrasilFil: Barbeito, Rafael Alonso. Facultad de Medicina; ArgentinaFil: Abdelhay, Eliana. Instituto Nacional de Cáncer; BrasilFil: Artagaveytia, Nora. Hospital de Clínicas Manuel Quintela. Universidad de la República; UruguayFil: Daneri-Navarro, Adrian. Universidad de Guadalajara; MéxicoFil: Llera, Andrea S. CONICET. Fundación Instituto Leloir; ArgentinaFil: Müller, Bettina. Instituto Nacional del Cáncer; ArgentinaFil: Podhajcer, Osvaldo L. CONICET. Fundación Instituto Leloir; ArgentinaFil: Velazquez, Carlos. Universidad de Sonora; MéxicoFil: Alcoba, Elsa. Hospital Municipal de Oncología María Curie; ArgentinaFil: Alonso, Isabel. Centro Hospitalario Pereira Rossell; ArgentinaFil: Bravo, Alicia I. Hospital Regional de Agudos Eva Perón; ArgentinaFil: Camejo, Natalia. Hospital de Clínicas Manuel Quintela. Universidad de la República; UruguayFil: Carraro, Dirce Maria. AC Camargo Cancer Center; BrasilFil: Castro, Mónica. Instituto de Oncología Angel Roffo; ArgentinaFil: Cataldi, Sandra. Instituto Nacional de Cáncer; UruguayFil: Cayota, Alfonso. Institut Pasteur de Montevideo; UruguayFil: Cerda, Mauricio. Universidad de Chile; ChileFil: Colombo, Alicia. Universidad de Chile; ChileFil: Crocamo, Susanne. Instituto Nacional de Cáncer; BrasilFil: Del Toro-Arreola, Alicia. Universidad de Guadalajara; MéxicoFil: Delgadillo-Cristerna, Raul. Hospital de Especialidades. CMNO-IMSS; MéxicoFil: Delgado, Lucia. Hospital de Clínicas Manuel Quintela; UruguayFil: Breitenbach, Marisa Dreyer. Universidade do Estado do Rio de Janeiro; BrasilFil: Fernández, Elmer. Universidad Católica de Córdoba. CONICET. Centro de Investigaciones en Bioquímica Clínica e Inmunologia; ArgentinaFil: Fernández, Jorge. Instituto de Salud Pública; ChileFil: Fernández, Wanda. Hospital San Borja Arriarán; ChileFil: Franco-Topete, Ramon A. OPD Hospital Civil de Guadalajara. Universidad de Guadalajara; MéxicoFil: Gaete, Fancy. Hospital Luis Tisne; ChileFil: Gómez, Jorge. Texas A&M University; Estados UnidosFil: Gonzalez-Ramirez, Leivy P. Universidad de Guadalajara; MéxicoFil: Guerrero, Marisol. Hospital San José; ChileFil: Gutierrez-Rubio, Susan A. Universidad de Guadalajara; MéxicoFil: Jalfin, Beatriz. Hospital Regional de Agudos Eva Perón; ArgentinaFil: Lopez-Vazquez, Alejandra. Universidad de Sonora; MéxicoFil: Loria, Dora. Instituto de Oncología Angel Roffo; ArgentinaFil: Míguez, Silvia. Hospital Municipal de Oncología María Curie; ArgentinaFil: Moran-Mendoza, Andres de J. Hospital de Gineco-Obstetricia CMNO-IMSS; MéxicoFil: Morgan-Villela, Gilberto. Hospital de Especialidades. CMNO-IMSS; MéxicoFil: Mussetti, Carina. Registro Nacional de Cancer; UruguayFil: Nagai, Maria Aparecida. Instituto de Câncer de São Paulo; BrasilFil: Oceguera-Villanueva, Antonio. Instituto Jalisciense de Cancerologia; MéxicoFil: Reis, Rui M. Hospital de Câncer de Barretos; BrasilFil: Retamales, Javier. Grupo Oncológico Cooperativo Chileno de Investigación; ChileFil: Rodriguez, Robinson. Hospital Central de las Fuerzas Armadas; UruguayFil: Rosales, Cristina, Hospital Municipal de Oncología María Curie; ArgentinaFil: Salas-Gonzalez, Efrain. Hospital San José; ChileFil: Segovia, Laura. Hospital Barros Luco Trudeau; ChileFil: Sendoya, Juan M. CONICET. Fundación Instituto Leloir,; ArgentinaFil: Silva-Garcia, Aida A. OPD Hospital Civil de Guadalajara. Universidad de Guadalajara; MéxicoFil: Viña, Stella. Instituto de Oncología Angel Roffo; ArgentinaFil: Zagame, Livia. Instituto Jalisciense de Cancerologia; MéxicoFil: Jones, Beth. Yale University. Yale School of Public Health; Estados UnidosFil: Szklo, Moysés. Johns Hopkins University. Johns Hopkins Bloomberg School of Public Health; Estados Unido