Bradley Hamilton

All Publications

• Cell-line and Antibody Epitope-dependent Expression Models Inform Selection of AAV-GNE Epimerase/Kinase Transgenes to Treat GNE Myopathy Patil, R., Hamilton, B., Day, J. CELL PRESS. 2025

  View details for Web of Science ID 001521601100226

• Extracellular Vesicles Reduce Type I Interferon Responses to AAV Gene Therapy in a Human Model of Innate Immunity Hamilton, B., Cheng, M., Patil, R., Wright, F., Crain, A., Day, J., Maguire, C. CELL PRESS. 2025

  View details for Web of Science ID 001587763700017

• Innate Immune Avoidance by Extracellular Vesicle Associated AAV (EV-AAV) Hamilton, B. A., de la Cruz, D., Patil, R., Cheng, M., Wright, J., Maguire, C. A. CELL PRESS. 2024: 727

  View details for Web of Science ID 001332783402482

• A Strategy to Select Minimally Immunogenic Candidate AAV Vectors to Treat GNE Myopathy Hamilton, B. A., Patil, R., Ruland, B., Day, J., Wright, J. CELL PRESS. 2024: 726-727

  View details for Web of Science ID 001332783402481

• A Biosynthetic Approach for CpG Methylation of AAV Expression Cassettes Huellen, F., Patil, R., Hamilton, B. A., Wright, J. CELL PRESS. 2023: 113-114

  View details for Web of Science ID 001045144200211

• A Human PBMC Assay of Type 1 Interferon Responses to Closely Related AAV Vectors Hamilton, B. A., Patil, R., Kim, S., Day, J. W., Wright, J. CELL PRESS. 2023: 76

  View details for Web of Science ID 001045144200141

• Methylation of CpG Dinucleotides in AAV Vector Plasmids Reduces TLR9 Immune Activation Patil, R., Hamilton, B., Perriman, R., Wright, J. CELL PRESS. 2022: 66-67

  View details for Web of Science ID 000794043700125

• ITRS FROM AAV SEROTYPES DIFFERENTIALLY STIMULATE INNATE IMMUNE SIGNALING THROUGH TLR9 Hamilton, B., Patil, R., Wright, F. ELSEVIER SCIENCE INC. 2022: S92

  View details for Web of Science ID 000890643400145

• Transduction of Pig Small Airway Epithelial Cells and Distal Lung Progenitor Cells by AAV4 CELLS Chen, O. G., Mather, S. E., Brommel, C. M., Hamilton, B. A., Ehler, A., Villacreses, R., Girgis, R. E., Abou Alaiwa, M., Stoltz, D. A., Zabner, J., Li, X. 2021; 10 (5)

  Abstract

  Cystic fibrosis (CF) is caused by genetic mutations of the CF transmembrane conductance regulator (CFTR), leading to disrupted transport of Cl- and bicarbonate and CF lung disease featuring bacterial colonization and chronic infection in conducting airways. CF pigs engineered by mutating CFTR develop lung disease that mimics human CF, and are well-suited for investigating CF lung disease therapeutics. Clinical data suggest small airways play a key role in the early pathogenesis of CF lung disease, but few preclinical studies have focused on small airways. Efficient targeted delivery of CFTR cDNA to small airway epithelium may correct the CFTR defect and prevent lung infections. Adeno-associated virus 4 (AAV4) is a natural AAV serotype and a safe vector with lower immunogenicity than other gene therapy vectors such as adenovirus. Our analysis of AAV natural serotypes using cultured primary pig airway epithelia showed that AAV4 has high tropism for airway epithelia and higher transduction efficiency for small airways compared with large airways. AAV4 mediated the delivery of CFTR, and corrected Cl- transport in cultured primary small airway epithelia from CF pigs. Moreover, AAV4 was superior to all other natural AAV serotypes in transducing ITGα6β4+ pig distal lung progenitor cells. In addition, AAV4 encoding eGFP can infect pig distal lung epithelia in vivo. This study demonstrates AAV4 tropism in small airway progenitor cells, which it efficiently transduces. AAV4 offers a novel tool for mechanistical study of the role of small airway in CF lung pathogenesis in a preclinical large animal model.

  View details for DOI 10.3390/cells10051014

  View details for Web of Science ID 000654729400001

  View details for PubMedID 33923029

  View details for PubMedCentralID PMC8145967

• Challenges Posed by Immune Responses to AAV Vectors: Addressing Root Causes. Frontiers in immunology Hamilton, B. A., Wright, J. F. 2021; 12: 675897

  Abstract

  Host immune responses that limit durable therapeutic gene expression and cause clinically significant inflammation remain a major barrier to broadly successful development of adeno-associated virus (AAV)-based human gene therapies. In this article, mechanisms of humoral and cellular immune responses to the viral vector are discussed. A perspective is provided that removal of pathogen-associated molecular patterns in AAV vector genomes to prevent the generation of innate immune danger signals following administration is a key strategy to overcome immunological barriers.

  View details for DOI 10.3389/fimmu.2021.675897

  View details for PubMedID 34084173

• Polarized AAVR expression determines infectivity by AAV gene therapy vectors GENE THERAPY Hamilton, B. A., Li, X., Pezzulo, A. A., Abou Alaiwa, M. H., Zabner, J. 2019; 26 (6): 240-249

  Abstract

  Adeno-associated virus (AAV) has been investigated to transfer the cystic fibrosis transmembrane conductance regulator (CFTR) to airways. Inhaled AAV2-CFTR in people with cystic fibrosis (CF) is safe, but inefficient. In vitro, AAV2 transduction of human airway epithelia on the apical (luminal) side is inefficient, but efficient basolaterally. We previously selected AAV2.5T, a novel capsid that apically transduces CF human airway epithelia and efficiently restores CFTR function. We hypothesize the AAV receptor (AAVR) is basolaterally localized, and that AAV2.5T utilizes an alternative apical receptor. We found AAVR in human airway epithelia by western blot and RNA-Seq analyses. Using immunocytochemistry we did not find endogenous AAVR at membranes but overexpression localized AAVR to the basolateral membrane, where it preferentially increased transduction. Anti-AAVR antibodies blocked transduction by AAV2 from the basolateral side but not AAV2.5T from the apical side, suggesting a unique apical receptor. Finally, we found infection by AAV2 but not AAV2.5T was blocked by CRISPR knockout of AAVR in cell lines. Our data suggest the absence of apical AAVR is rate limiting for AAV2, and efficient transduction by AAV2.5T is accomplished using an AAVR independent pathway. Our findings inform the development of gene therapy for CF, and AAV vectors in general.

  View details for DOI 10.1038/s41434-019-0078-3

  View details for Web of Science ID 000472146900003

  View details for PubMedID 30962536

  View details for PubMedCentralID PMC6588428

• EGF regulation of proximal tubule cell proliferation and VEGF-A secretion PHYSIOLOGICAL REPORTS Zepeda-Orozco, D., Wen, H. M., Hamilton, B. A., Raikwar, N. S., Thomas, C. P. 2017; 5 (18)

  Abstract

  Proximal tubule cell (PTC) proliferation is critical for tubular regeneration and recovery from acute kidney injury. Epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF-A) are important for the maintenance of tubulointerstitial integrity and can stimulate PTC proliferation. We utilized HK-2 cells, an immortalized human PTC line, to characterize the EGF-dependent regulation of VEGF-A secretion and proliferation in PTCs. We demonstrate that EGF stimulates VEGF-A secretion via the EGF receptor (EGFR) and stimulates cell proliferation via activation of the VEGF receptor, VEGFR-2. EGFR activation promotes MAPK (ERK1/2) activation and HIF-1α expression, which are required for basal and EGF-stimulated VEGF-A secretion. EGF also stimulates the phosphorylation of P70S6 kinase (P70S6K), the downstream target of mTORC1. Rapamycin decreased basal and EGF stimulated HIF-1α and enhanced MAPK (ERK1/2) activation, while MAPK (ERK/12) inhibition downregulated HIF-1α expression and the phosphorylation of p70S6K. EGF stimulation of p70S6K was also independent of p-AKT Inhibition of the mTORC1 pathway with rapamycin abolished phosphorylation of p70S6K but had no effect on VEGF-A secretion, indicating that EGF-stimulated VEGF-A secretion did not require mTORC1 pathway activation. We demonstrate evidence of a complex crosstalk between the MAPK/ERK and mTORC1 pathways, wherein MAPK (ERK1/2) activation stimulates p-P70S6K, while p-P70S6K activation seems to inhibit MAPK (ERK1/2) in EGF-treated HK-2 cells. Our results suggest that EGF stimulates MAPK (ERK1/2) in HK-2 cells, which in turn increases HIF-1α expression and VEGF-A secretion, indicating that VEGF-A mediates EGF-stimulated cell proliferation as an autocrine proximal tubular epithelial cell growth factor.

  View details for DOI 10.14814/phy2.13453

  View details for Web of Science ID 000417428200013

  View details for PubMedID 28963126

  View details for PubMedCentralID PMC5617933