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Precision Cas9 Genome Editing In Vivo With All-In-One, Self-Targeting Aav Vectors [Preprint], Raed Ibraheim, Phillip W. L. Tai, Aamir Mir, Nida Javeed, Jiaming Wang, Tomas Rodriguez, Samantha J. Nelson, Eraj Khokhar, Esther Mintzer, Stacy Maitland​, Yueying Cao, Guangping Gao, Erik J. Sontheimer 2020 University of Massachusetts Medical School

Precision Cas9 Genome Editing In Vivo With All-In-One, Self-Targeting Aav Vectors [Preprint], Raed Ibraheim, Phillip W. L. Tai, Aamir Mir, Nida Javeed, Jiaming Wang, Tomas Rodriguez, Samantha J. Nelson, Eraj Khokhar, Esther Mintzer, Stacy Maitland​, Yueying Cao, Guangping Gao, Erik J. Sontheimer

University of Massachusetts Medical School Faculty Publications

Adeno-associated virus (AAV) vectors are important delivery platforms for therapeutic genome editing but are severely constrained by cargo limits, especially for large effectors like Cas9s. Simultaneous delivery of multiple vectors can limit dose and efficacy and increase safety risks. The use of compact effectors has enabled single-AAV delivery of Cas9s with 1-3 guides for edits that use end-joining repair pathways, but many precise edits that correct disease-causing mutations in vivo require homology-directed repair (HDR) templates. Here, we describe single-vector, ~4.8-kb AAV platforms that express Nme2Cas9 and either two sgRNAs to produce segmental deletions, or a single sgRNA with an ...


Astrocyte- And Neuron-Derived Cxcl1 Drives Neutrophil Transmigration And Blood-Brain Barrier Permeability In Viral Encephalitis, Benedict D. Michael, Laura Bricio-Moreno, Elizabeth W. Sorensen, Yoshishige Miyabe, Jeffrey Lian, Tom Solomon, Evelyn A. Kurt-Jones, Andrew D. Luster 2020 Harvard Medical School

Astrocyte- And Neuron-Derived Cxcl1 Drives Neutrophil Transmigration And Blood-Brain Barrier Permeability In Viral Encephalitis, Benedict D. Michael, Laura Bricio-Moreno, Elizabeth W. Sorensen, Yoshishige Miyabe, Jeffrey Lian, Tom Solomon, Evelyn A. Kurt-Jones, Andrew D. Luster

Open Access Articles

Herpes simplex virus (HSV)-1 encephalitis has significant morbidity partly because of an over-exuberant immune response characterized by leukocyte infiltration into the brain and increased blood-brain barrier (BBB) permeability. Determining the role of specific leukocyte subsets and the factors that mediate their recruitment into the brain is critical to developing targeted immune therapies. In a murine model, we find that the chemokines CXCL1 and CCL2 are induced in the brain following HSV-1 infection. Ccr2 (CCL2 receptor)-deficient mice have reduced monocyte recruitment, uncontrolled viral replication, and increased morbidity. Contrastingly, Cxcr2 (CXCL1 receptor)-deficient mice exhibit markedly reduced neutrophil recruitment, BBB ...


Molecular Mechanisms Lead To Sex-Specific Covid-19 Prognosis And Targeted Therapies, Thushara Galbadage, Brent M. Peterson, Jeffrey S. Wang, Avishka Jayasekara, Danny A. Ramirez, Joseph Awada, John P. Walsh, Richard S. Gunasekera 2020 Biola University

Molecular Mechanisms Lead To Sex-Specific Covid-19 Prognosis And Targeted Therapies, Thushara Galbadage, Brent M. Peterson, Jeffrey S. Wang, Avishka Jayasekara, Danny A. Ramirez, Joseph Awada, John P. Walsh, Richard S. Gunasekera

Faculty Articles & Research

Clinical and epidemiological studies have identified male sex as an important risk factor for COVID-19 clinical outcomes and mortality. This raises the question as to how this risk factor can be addressed in the prognosis, clinical management, and the treatment of patients with Coronavirus disease 2019 (COVID-19). Currently, there are no guidelines or protocols to help alter the course of sex-specific COVID-19 prognosis, especially in severe disease presentations. This is partly due to the lack of research studies characterizing the differences in male vs. female host response to the severe acute respiratory syndrome Coronavirus-2 (SARS-CoV-2) infection and a lack of ...


Delayed Interventions, Low Compliance, And Health Disparities Amplified The Early Spread Of Covid-19, Aliea M. Jalali, Sumaia G. Khoury, JongWon See, Alexis M. Gulsvig, Brent M. Peterson, Richard S. Gunasekera, Gentian Buzi, Jason Wilson, Thushara Galbadage 2020 Biola University

Delayed Interventions, Low Compliance, And Health Disparities Amplified The Early Spread Of Covid-19, Aliea M. Jalali, Sumaia G. Khoury, Jongwon See, Alexis M. Gulsvig, Brent M. Peterson, Richard S. Gunasekera, Gentian Buzi, Jason Wilson, Thushara Galbadage

Faculty Articles & Research

The United States (US) public health interventions were rigorous and rapid, yet failed to arrest the spread of the Coronavirus Disease 2019 (COVID-19) pandemic as infections spread throughout the US. Many factors have contributed to the spread of COVID-19, and the success of public health interventions depends on the level of community adherence to preventative measures. Public health professionals must also understand regional demographic variation in health disparities and determinants to target interventions more effectively. In this study, a systematic evaluation of three significant interventions employed in the US, and their effectiveness in slowing the early spread of COVID-19 was ...


Structural Analysis Of Potent Hybrid Hiv-1 Protease Inhibitors Containing Bis-Tetrahydrofuran In A Pseudo-Symmetric Dipeptide Isostere, Linah Rusere, Gordon J. Lockbaum, Mina Henes, Sook-Kyung Lee, Ean Spielvogel, Desaboini Nageswara Rao, Klajdi Kosovrasti, Ellen A. Nalivaika, Ronald Swanstrom, Nese Kurt Yilmaz, Celia A. Schiffer, Akbar Ali 2020 University of Massachusetts Medical School

Structural Analysis Of Potent Hybrid Hiv-1 Protease Inhibitors Containing Bis-Tetrahydrofuran In A Pseudo-Symmetric Dipeptide Isostere, Linah Rusere, Gordon J. Lockbaum, Mina Henes, Sook-Kyung Lee, Ean Spielvogel, Desaboini Nageswara Rao, Klajdi Kosovrasti, Ellen A. Nalivaika, Ronald Swanstrom, Nese Kurt Yilmaz, Celia A. Schiffer, Akbar Ali

University of Massachusetts Medical School Faculty Publications

The design, synthesis, and X-ray structural analysis of hybrid HIV-1 protease inhibitors (PIs) containing bis-tetrahydrofuran (bis-THF) in a pseudo-C2-symmetric dipeptide isostere are described. A series of PIs were synthesized by incorporating bis-THF of darunavir on either side of the Phe-Phe isostere of lopinavir in combination with hydrophobic amino acids on the opposite P2/P2' position. Structure-activity relationship studies indicated that the bis-THF moiety can be attached at either the P2 or P2' position without significantly affecting potency. However, the group on the opposite P2/P2' position had a dramatic effect on potency depending on the size and shape of the ...


Acidic Ph Triggers Lipid Mixing Mediated By Lassa Virus Gp, Uriel Bulow, Ramesh Govindan, James B. Munro 2020 Tufts University

Acidic Ph Triggers Lipid Mixing Mediated By Lassa Virus Gp, Uriel Bulow, Ramesh Govindan, James B. Munro

Open Access Articles

Lassa virus (LASV) is the causative agent of Lassa hemorrhagic fever, a lethal disease endemic to Western Africa. LASV entry is mediated by the viral envelope glycoprotein (GP), a class I membrane fusogen and the sole viral surface antigen. Previous studies have identified components of the LASV entry pathway, including several cellular receptors and the requirement of endosomal acidification for infection. Here, we first demonstrate that incubation at a physiological temperature and pH consistent with the late endosome is sufficient to render pseudovirions, bearing LASV GP, non-infectious. Antibody binding indicates that this loss of infectivity is due to a conformational ...


Mrna Stem-Loops Can Pause The Ribosome By Hindering A-Site Trna Binding, Chen Bao, Sarah Loerch, Clarence Ling, Andrei A. Korostelev, Nikolaus Grigorieff, Dmitri N. Ermolenko 2020 University of Rochester

Mrna Stem-Loops Can Pause The Ribosome By Hindering A-Site Trna Binding, Chen Bao, Sarah Loerch, Clarence Ling, Andrei A. Korostelev, Nikolaus Grigorieff, Dmitri N. Ermolenko

Open Access Articles

Although the elongating ribosome is an efficient helicase, certain mRNA stem-loop structures are known to impede ribosome movement along mRNA and stimulate programmed ribosome frameshifting via mechanisms that are not well understood. Using biochemical and single-molecule Forster resonance energy transfer (smFRET) experiments, we studied how frameshift-inducing stem-loops from E. coli dnaX mRNA and the gag-pol transcript of Human Immunodeficiency Virus (HIV) perturb translation elongation. We find that upon encountering the ribosome, the stem-loops strongly inhibit A-site tRNA binding and ribosome intersubunit rotation that accompanies translation elongation. Electron cryo-microscopy (cryo-EM) reveals that the HIV stem-loop docks into the A site of ...


Gp41-Targeted Antibodies Restore Infectivity Of A Fusion-Deficient Hiv-1 Envelope Glycoprotein, Vinita R. Joshi, Ruchi M. Newman, Melissa L. Pack, Karen A. Power, James B. Munro, Ken Okawa, Navid Madani, Joseph G. Sodroski, Aaron G. Schmidt, Todd M. Allen 2020 Harvard Medical School

Gp41-Targeted Antibodies Restore Infectivity Of A Fusion-Deficient Hiv-1 Envelope Glycoprotein, Vinita R. Joshi, Ruchi M. Newman, Melissa L. Pack, Karen A. Power, James B. Munro, Ken Okawa, Navid Madani, Joseph G. Sodroski, Aaron G. Schmidt, Todd M. Allen

Open Access Articles

The HIV-1 envelope glycoprotein (Env) mediates viral entry via conformational changes associated with binding the cell surface receptor (CD4) and coreceptor (CCR5/CXCR4), resulting in subsequent fusion of the viral and cellular membranes. While the gp120 Env surface subunit has been extensively studied for its role in viral entry and evasion of the host immune response, the gp41 transmembrane glycoprotein and its role in natural infection are less well characterized. Here, we identified a primary HIV-1 Env variant that consistently supports >300% increased viral infectivity in the presence of autologous or heterologous HIV-positive plasma. However, in the absence of HIV-positive ...


Characterization Of A Plasmid-Based Dna Vaccine For Simian Immunodeficiency Virus, Priya Sanipara 2020 University of Mississippi Main Campus

Characterization Of A Plasmid-Based Dna Vaccine For Simian Immunodeficiency Virus, Priya Sanipara

Honors Theses

Described as one of the world’s worst pandemics, HIV (Human Immunodeficiency Virus) infects millions of people each year and is the cause for AIDS (Acquired Immunodeficiency Syndrome). Despite the development of vaccines for numerous infectious diseases such as polio, small pox, and influenza, a vaccine for HIV remains elusive due to the virus’s high mutation rate and ability to evade the immune system. HIV causes depletion of CD4+ lymphocytes, resulting in a weakened immune system. However, the development of a plasmid-based DNA vaccine approach may help revolutionize vaccine development for HIV due to its ability to confer cellular ...


Life During A Pandemic, Srinivasan Ragothaman 2020 Srini.Ragothaman@usd.edu

Life During A Pandemic, Srinivasan Ragothaman

Creative Work

This is a short poem (6 stanzas) about the current pandemic and the trials and tribulations people are going through. This describes the heroic efforts by the doctors, nurses, and other frontline warriors in tackling the virus related illnesses and pays a rich tribute to all heroes from every walk of life who are in the front fighting this disease. It hopes for a plan, a vaccine, and a cure soon. This poem also describes the switchover to remote instruction at a reginal University, student and faculty adjustments to the pandemic and pokes gentle fun at the opportunity to watch ...


Smrt Sequencing Of Paramecium Bursaria Chlorella Virus-1 Reveals Diverse Methylation Stability In Adenines Targeted By Restriction Modification Systems, Samantha R. Coy, Eric R. Gann, Spiridon E. Papoulis, Michael E. Holder, Nadim J. Ajami, Joseph F. Petrosino, Erik R. Zinser, James L. Van Etten, Steven W. Wilhelm 2020 University of Tennessee, Knoxville & Rice University

Smrt Sequencing Of Paramecium Bursaria Chlorella Virus-1 Reveals Diverse Methylation Stability In Adenines Targeted By Restriction Modification Systems, Samantha R. Coy, Eric R. Gann, Spiridon E. Papoulis, Michael E. Holder, Nadim J. Ajami, Joseph F. Petrosino, Erik R. Zinser, James L. Van Etten, Steven W. Wilhelm

James Van Etten Publications

Chloroviruses (family Phycodnaviridae) infect eukaryotic, freshwater, unicellular green algae. A unique feature of these viruses is an abundance of DNA methyltransferases, with isolates dedicating up to 4.5% of their protein coding potential to these genes. This diversity highlights just one of the long-standing values of the chlorovirus model system; where group-wide epigenomic characterization might begin to elucidate the function(s) of DNA methylation in large dsDNA viruses. We characterized DNA modifications in the prototype chlorovirus, PBCV-1, using single-molecule real time (SMRT) sequencing (aka PacBio). Results were compared to total available sites predicted in silico based on DNA sequence alone ...


How Immune T-Cell Augmentation Can Help Prevent Covid-19: A Possible Nutritional Solution Using Ketogenic Lifestyle, Ravi k. Kamepalli MD,FIDSA,CWSP 2020 Regional Infectious diseases and infusion ctr,Inc

How Immune T-Cell Augmentation Can Help Prevent Covid-19: A Possible Nutritional Solution Using Ketogenic Lifestyle, Ravi K. Kamepalli Md,Fidsa,Cwsp

The University of Louisville Journal of Respiratory Infections

No abstract provided.


Impact Of The Affordable Care Act On Referral To Care For People Living With Hiv In Appalachia, Cameron A. Wade, Timothy N. Crawford, Nicole Leedy, Alice C. Thornton 2020 University of Kentucky College of Medicine

Impact Of The Affordable Care Act On Referral To Care For People Living With Hiv In Appalachia, Cameron A. Wade, Timothy N. Crawford, Nicole Leedy, Alice C. Thornton

Journal of Appalachian Health

Introduction: The Affordable Care Act (ACA) enacted on March 23, 2010 significantly impacted access to healthcare for people living with HIV (PLWH). Expansion of care was accomplished in three areas: eliminating exclusions for pre-existing conditions, elimination of lifetime caps on healthcare expenditures, and expansion of Medicaid eligibility.

Purpose: This study evaluated the impact of state implementation of the ACA Medicaid expansion on referral to HIV care at a Ryan White federally funded clinic in Kentucky (University of Kentucky Bluegrass Care Clinic [UK BCC]).

Methods: Retrospective chart review of all newly enrolled patients at the UK BCC between March 2010 and ...


Trim34 Restricts Hiv-1 And Siv Capsids In A Trim5alpha-Dependent Manner, Molly Ohainle, Kyusik Kim, Sevnur Komurlu Keceli, Abby Felton, Ed Campbell, Jeremy Luban, Michael Emerman 2020 Fred Hutchinson Cancer Research Center

Trim34 Restricts Hiv-1 And Siv Capsids In A Trim5alpha-Dependent Manner, Molly Ohainle, Kyusik Kim, Sevnur Komurlu Keceli, Abby Felton, Ed Campbell, Jeremy Luban, Michael Emerman

Open Access Articles

The HIV-1 capsid protein makes up the core of the virion and plays a critical role in early steps of HIV replication. Due to its exposure in the cytoplasm after entry, HIV capsid is a target for host cell factors that act directly to block infection such as TRIM5alpha and MxB. Several host proteins also play a role in facilitating infection, including in the protection of HIV-1 capsid from recognition by host cell restriction factors. Through an unbiased screening approach, called HIV-CRISPR, we show that the CPSF6-binding deficient, N74D HIV-1 capsid mutant is sensitive to restriction mediated by human TRIM34 ...


Declines In Lung Function After Antiretroviral Therapy Initiation In Adults With Human Immunodeficiency Virus And Tuberculosis: A Potential Manifestation Of Respiratory Immune Reconstitution Inflammatory Syndrome, Sara C. Auld, Pholo Maenetje, Shruthi Ravimohan, Drew Weissman, Itai Ncube, Mandla Mlotshwa, Nelly Ratsela, William Chase, Mboyo-Di-Tamba Vangu, Robert Wallis, Gavin Churchyard, Hardy Kornfeld, Gregory P. Bisson 2020 Emory University

Declines In Lung Function After Antiretroviral Therapy Initiation In Adults With Human Immunodeficiency Virus And Tuberculosis: A Potential Manifestation Of Respiratory Immune Reconstitution Inflammatory Syndrome, Sara C. Auld, Pholo Maenetje, Shruthi Ravimohan, Drew Weissman, Itai Ncube, Mandla Mlotshwa, Nelly Ratsela, William Chase, Mboyo-Di-Tamba Vangu, Robert Wallis, Gavin Churchyard, Hardy Kornfeld, Gregory P. Bisson

Open Access Articles

End-organ impairment has received relatively little research attention as a possible manifestation of tuberculosis immune reconstitution inflammatory syndrome (TB-IRIS). In this prospective cohort study, one-half of adults with human immunodeficiency virus and pulmonary tuberculosis experienced meaningful declines in lung function on antiretroviral therapy, suggesting a role for lung function in TB-IRIS definitions.


Welcome To The Peer Covid-19 Seminar, Patricia A. McQuilkin 2020 University of Massachusetts Medical School

Welcome To The Peer Covid-19 Seminar, Patricia A. Mcquilkin

PEER Liberia Project

This presentation welcomes attendees and provides the agenda for the online PEER Liberia COVID-19 Seminar.


Epidemic To Endemic Or Pandemic Infectious Diseases: Commonalities And Distinctions Between Malaria, Ebola And Covid19, Ann M. Moormann 2020 University of Massachusetts Medical School

Epidemic To Endemic Or Pandemic Infectious Diseases: Commonalities And Distinctions Between Malaria, Ebola And Covid19, Ann M. Moormann

PEER Liberia Project

This presentation provides an overview of epidemics by discussing malaria, Ebola and COVID-19: definition of terms; examples of diseases, transmission and health impact; diagnostics versus presumptive diagnosis –how to make strategic decisions with limited resources and specific context; how strategies are modified by what we learn -- how fast can we adapt; control, elimination, eradication -- modeling the end game.


On The Inadequacy Of Species Distribution Models For Modelling The Spread Of Sars-Cov-2: Response To Araújo And Naimi, Joseph D. Chipperfield, Blas M. Benito, Robert B. O'Hara, Richard J. Telford, Colin J. Carlson 2020 Norwegian Institute for Nature Research

On The Inadequacy Of Species Distribution Models For Modelling The Spread Of Sars-Cov-2: Response To Araújo And Naimi, Joseph D. Chipperfield, Blas M. Benito, Robert B. O'Hara, Richard J. Telford, Colin J. Carlson

Public Health Resources

The ongoing pandemic of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is causing significant damage to public health and economic livelihoods, and is putting significant strains on healthcare services globally. This unfolding emergency has prompted the preparation and dissemination of the article “Spread of SARS-CoV-2 Coronavirus likely to be constrained by climate” by Araújo and Naimi (2020). The authors present the results of an ensemble forecast made from a suite of species distribution models (SDMs), where they attempt to predict the suitability of the climate for the spread of SARS-CoV-2 over the coming months. They argue that climate is ...


Comparisons Of Antibody Populations In Different Pre-Fusion F Vlp-Immunized Cotton Rat Dams And Their Offspring, Lori McGinnes Cullen, Marina S. Boukhvalova, Jorge C. G. Blanco, Trudy G. Morrison 2020 University of Massachusetts Medical School

Comparisons Of Antibody Populations In Different Pre-Fusion F Vlp-Immunized Cotton Rat Dams And Their Offspring, Lori Mcginnes Cullen, Marina S. Boukhvalova, Jorge C. G. Blanco, Trudy G. Morrison

Open Access Articles

Respiratory syncytial virus (RSV) infection poses a significant risk for infants. Since the direct vaccination of infants is problematic, maternal vaccination may provide a safer, more effective approach to their protection. In the cotton rat (CR) model, we have compared the immunization of pregnant CR dams with virus-like particles assembled with the prototype mutation stabilized pre-fusion F protein, DS-Cav1, as well two alternative mutation stabilized pre-fusion proteins (UC-2 F, UC-3 F) and showed that the alternative pre-fusion F VLPs protected the offspring of immunized dams significantly better than DS-Cav1 F VLPs (Blanco, et al. J. Virol. 93: e00914). Here, we ...


Epstein-Barr Virus Epitope-Major Histocompatibility Complex Interaction Combined With Convergent Recombination Drives Selection Of Diverse T Cell Receptor Alpha And Beta Repertoires, Anna Gil, Larisa Kamga, Ramakanth Chirravuri-Venkata, Nuray Aslan, Fransenio G. Clark, Dario Ghersi, Katherine Luzuriaga, Liisa K. Selin 2020 University of Massachusetts Medical School

Epstein-Barr Virus Epitope-Major Histocompatibility Complex Interaction Combined With Convergent Recombination Drives Selection Of Diverse T Cell Receptor Alpha And Beta Repertoires, Anna Gil, Larisa Kamga, Ramakanth Chirravuri-Venkata, Nuray Aslan, Fransenio G. Clark, Dario Ghersi, Katherine Luzuriaga, Liisa K. Selin

Open Access Articles

Recognition modes of individual T cell receptors (TCRs) are well studied, but factors driving the selection of TCR repertoires from primary through persistent human virus infections are less well understood. Using deep sequencing, we demonstrate a high degree of diversity of Epstein-Barr virus (EBV)-specific clonotypes in acute infectious mononucleosis (AIM). Only 9% of unique clonotypes detected in AIM persisted into convalescence; the majority (91%) of unique clonotypes detected in AIM were not detected in convalescence and were seeming replaced by equally diverse "de novo" clonotypes. The persistent clonotypes had a greater probability of being generated than nonpersistent clonotypes due ...


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