A5024981373- Virus-based gene therapy research
- Viral Infectious Diseases and Gene Expression in Insects
- Protease and Inhibitor Mechanisms
- CRISPR and Genetic Engineering
- Viral Infections and Immunology Research
- Animal Genetics and Reproduction
- Neonatal Respiratory Health Research
- RNA Interference and Gene Delivery
- Peptidase Inhibition and Analysis
- Neurogenetic and Muscular Disorders Research
- CAR-T cell therapy research
- Muscle Physiology and Disorders
- Viral gastroenteritis research and epidemiology
- Amyotrophic Lateral Sclerosis Research
- RNA Research and Splicing
- Blood Coagulation and Thrombosis Mechanisms
- RNA modifications and cancer
- Congenital Diaphragmatic Hernia Studies
- Veterinary Equine Medical Research
- Veterinary Medicine and Surgery
- Signaling Pathways in Disease
- Tissue Engineering and Regenerative Medicine
- Dermatological diseases and infestations
- Odor and Emission Control Technologies
- Adenosine and Purinergic Signaling
University of Massachusetts Chan Medical School
2016-2025
Tufts University
2011-2025
Gene Therapy Laboratory
2022
UMass Memorial Medical Center
2022
Cummings Veterinary Medical Center
2021
UMass Memorial Health Care
2016
New England Disabled Sports
2009-2010
Purdue University West Lafayette
2006
α-1 antitrypsin (AAT) deficiency can exhibit two pathologic states: a lung disease that is primarily due to the loss of AAT's antiprotease function, and liver resulting from toxic gain-of-function PiZ-AAT (Z-AAT) mutant protein. We have developed several recombinant adeno-associated virus (rAAV) vectors incorporate microRNA (miRNA) sequences targeting AAT gene while also driving expression miRNA-resistant wild-type AAT-PiM (M-AAT) gene, thus achieving concomitant Z-AAT knockdown in increased...
Abstract Physiological regulation of transgene expression is a major challenge in gene therapy. Onasemnogene abeparvovec (Zolgensma ® ) an approved adeno-associated virus (AAV) vector therapy for infants with spinal muscular atrophy (SMA), however, adverse events have been observed both animals and patients following treatment. The construct contains native human survival motor neuron 1 ( hSMN1 driven by strong, cytomegalovirus enhancer/chicken β-actin CMVen/CB promoter providing high,...
Organ regeneration in mammals is hypothesized to require a functional pool of stem or progenitor cells, but the role these cells lung unknown. Whereas postnatal alveolar tissue has been attributed type II epithelial (AECII), we reasoned that bronchioalveolar (BASCs) have potential contribute substantially this process. To test hypothesis, unilateral pneumonectomy (PNX) was performed on adult female C57/BL6 mice stimulate compensatory regrowth. The density BASCs and AECII, morphometric...
Significance Chronic obstructive pulmonary disease affects 10% of the worldwide population, and leading genetic cause is a disease, α-1 antitrypsin (AAT) deficiency. Humans have only one gene that codes for AAT protein, but mice up to six, which made it impossible decades create mouse model disease. Here we succeeded in creating this using CRISPR technology target all genes at once. Importantly, spontaneously develops lung recapitulates many aspects human We anticipate will be highly...
While aging leads to a reduction in the capacity for regeneration after pneumonectomy (PNX) most mammals, this biological phenomenon has not been characterized over lifetime of mice. We measured age-specific (3, 9, 24 month) effects PNX on physiology, morphometry, cell proliferation and apoptosis, global gene expression, lung fibroblast phenotype clonogenicity female C57BL6 The data show that only 3 month old mice were fully capable restoring volumes by day 7 total alveolar surface area 21...
The lung is a complex organ with various cell types having distinct roles. Antisense oligonucleotides (ASOs) have been studied in the lung, but it has challenging to determine their effectiveness each type due lack of appropriate analytical methods. We employed three approaches study silencing efficacy within different types. First, we used lineage markers identify flow cytometry, and simultaneously measured ASO-induced cell-surface proteins CD47 or CD98. Second, applied single-cell RNA...
Mechanical stress is an important modulator of lung morphogenesis, postnatal development, and compensatory regrowth. The effect mechanical on stem or progenitor cells unclear. We examined whether proliferative responses epithelial cells, including dually immunoreactive (CCSP proSP-C) (CCSP+/SP-C+) type II alveolar (ATII), are affected by physical factors found in the emphysematics, loss elastic recoil, reduced elastin content, destruction. Mice underwent single pneumonectomy (PNY) to...
Recombinant adeno-associated virus (rAAV) vectors are rapidly becoming the first choice for human gene therapy studies, as clinical efficacy has been demonstrated in several trials and proof-of-concept data have correction of many others. When moving into use under auspices an FDA Investigational New Drug (IND) application, it is necessary to demonstrate stability vector material various conditions storage, dilution, administration when used humans. Limited currently available literature...
With the US Food and Drug Administration (FDA) European Medicines Agency (EMA) approvals for Zolgensma, Luxturna, Glybera, recombinant adeno-associated viruses (rAAVs) are considered efficient tools gene transfer. However, studies in animals humans demonstrate that intramuscular (IM) AAV delivery can trigger immune responses to capsids and/or transgenes. IM of rAAV1 has also been described induce tolerance rAAV characterized by presence capsid-specific regulatory T cells (Tregs) periphery....
Immune responses to adeno-associated virus (AAV) capsids limit the therapeutic potential of AAV gene therapy. Herein, we model clinical immune by generating capsid-specific chimeric antigen receptor (AAV-CAR) T cells. We then modulate capsid with AAV-CAR regulatory cells (Tregs). Tregs in vitro display phenotypical Treg surface marker expression, and functional suppression effector cell proliferation cytotoxicity. In mouse models, mediated continued transgene expression from an immunogenic...
Classic maple syrup urine disease (MSUD) results from biallelic mutations in genes that encode the branched-chain α-ketoacid dehydrogenase E1 α ( BCKDHA ), β BCKDHB or dihydrolipoamide transacylase DBT ) subunits, which interact to form mitochondrial BCKDH complex decarboxylates ketoacid derivatives of leucine, isoleucine, and valine. MSUD is an inborn error metabolism characterized by recurrent life-threatening neurologic crises progressive brain injury can only be managed with exacting...
ABSTRACT AAV capsid serotypes isolated from nature have been widely used in gene delivery and therapy. Recently, more than 1,000 distinct capsids were identified human clinical samples by high-throughput, long-read DNA sequencing (Hsu HL et al. Nature Communications 2020). In this study, we tap into broad natural biodiversity of to develop liver-tropic capsids. We initially screened a subset variants derived AAV8 (n=159) for packaging efficiency. The top 30% these subjected barcoded vector...
Alpha-1 antitrypsin deficiency (AATD) is commonly caused by a G-to-A mutation in the SERPINA1 gene (the PiZ mutation). The mutant AAT protein sequestered hepatocytes, causing lung emphysema due to insufficient inhibit neutrophil elastase lung. Here we show that compact adenine base editor (ABE) with an evolved Cas9 nickase derived from Neisseria meningitidis (eNme2.C) can be packaged single AAV and correct mouse models of AATD. An all-in-one eNme2.C-TadA8e/guide 2 plasmid achieved...
Background: Renal biopsies are uncommonly performed in horses and little is known about their diagnostic utility associated complication rate. Objective: To describe the techniques, rate, risk factors, histopathology results; as well evaluate safety of renal biopsy horse. Animals: One hundred forty‐six from which 151 were obtained. Animals ranged age 48 hours to 30 years. Methods: Multicenter retrospective study, with participation 14 institutions (1983–2009). Results: was a similar rate...
Aging is a critical determinant of regenerative capacity in many organ systems, but it remains unresolved the lung. This study examines murine lung cell dynamics during age-dependent regeneration. Proliferation progenitor cells (EpCAMneg/Sca-1high mesenchymal stromal - LMSCs, EpCAMpos/Sca-1low epithelial cells, proSP-Cpos alveolar type II AECII, and CD31pos endothelial cells) was tracked to day 3 or 7 after pneumonectomy (PNX) SHAM surgery 3, 9, 17 month mice. In mice, post-PNX LMSC...
Abstract Adeno‐associated virus (AAV) is a DNA with small (∼4.7 kb) single‐stranded genome. It naturally replication‐defective parvovirus of the dependovirus group. Recombinant AAV (rAAV), for use as gene transfer vector, created by replacing viral rep and cap genes transgene interest along promoter polyadenylation sequences. Only inverted terminal repeats (ITRs) are required in cis replication packaging during production. The ITRs also necessary sufficient vector genome processing...
Alpha-1 antitrypsin deficiency (AATD) is characterized by both chronic lung disease due to loss of wild-type AAT (M-AAT) antiprotease function and liver toxicity from delayed secretion, polymerization, aggregation misfolded mutant (Z-AAT). The ideal gene therapy for AATD should therefore comprise endogenous Z-AAT suppression M-AAT overexpression. We designed a dual-function rAAV3B (df-rAAV3B) construct, which was effective at transducing hepatocytes, resulting in considerable decrease levels...
Abstract Measurements of total, respirable, and real-time airborne particulate were obtained in 12 horse barns summer winter. Respirable total concentrations measured gravimetrically; was with an aerosol photometer. Total (TP) ranged from nondetectable (ND) to 2.1 mg/m3 ND 1.2 for winter sampling, respectively. (RP) 0.2 0.7 measurements, The mean respirable fraction measurements 0.63 0.34, respectively Real-time (RTP) concentration significantly correlated TP pooled data but not RP winter,...
Abstract Adeno‐associated virus (AAV) is a small replication‐deficient DNA belonging to the Parvovirinae family. It has single‐stranded ∼4.7‐kb genome. Recombinant AAV (rAAV) created by replacing viral rep and cap genes with transgene of interest along promoter polyadenylation sequences. The short inverted terminal repeats must remain intact for replication packaging in production, as well vector genome processing persistence transduction process. capsid (serotype) determines tissue tropism...