Charles Gersbach

Positions:

John W. Strohbehn Distinguished Professor of Biomedical Engineering

Biomedical Engineering
Pratt School of Engineering

Professor of Biomedical Engineering

Biomedical Engineering
Pratt School of Engineering

Associate Professor of Surgery

Surgery, Surgical Sciences
School of Medicine

Associate Professor in Orthopaedic Surgery

Orthopaedic Surgery
School of Medicine

Associate Professor in Cell Biology

Cell Biology
School of Medicine

Associate of the Duke Initiative for Science & Society

Duke Science & Society
Institutes and Provost's Academic Units

Core Faculty in Innovation & Entrepreneurship

Duke Innovation & Entrepreneurship
Institutes and Provost's Academic Units

Member of the Duke Cancer Institute

Duke Cancer Institute
School of Medicine

Affiliate of the Duke Regeneration Center

Regeneration Next Initiative
School of Medicine

Education:

B.S. 2001

Georgia Institute of Technology

Ph.D. 2006

Georgia Institute of Technology

Grants:

Regulatory Mechanisms of CD4+ T Cell Differentiation

Administered By
Biostatistics & Bioinformatics
Awarded By
National Institutes of Health
Role
Co-Principal Investigator
Start Date
End Date

Regulatory Mechanisms of CD4+ T Cell Differentiation

Administered By
Integrative Genomics
Awarded By
National Institutes of Health
Role
Co-Principal Investigator
Start Date
End Date

Investigating Autophagy in GSD-Ia

Administered By
Pediatrics, Medical Genetics
Awarded By
National Institutes of Health
Role
Co Investigator
Start Date
End Date

Mechanotransduction in Meniscus Health and Repair

Administered By
Orthopaedics
Awarded By
National Institutes of Health
Role
Collaborator
Start Date
End Date

EFRI CEE : Engineering Technologies to Determine Causal Relationships Between Chromatin Structure and Gene Regulation

Administered By
Biomedical Engineering
Awarded By
National Science Foundation
Role
Principal Investigator
Start Date
End Date

Publications:

Robust, Durable Gene Activation In Vivo via mRNA-Encoded Activators.

Programmable control of gene expression via nuclease-null Cas9 fusion proteins has enabled the engineering of cellular behaviors. Here, both transcriptional and epigenetic gene activation via synthetic mRNA and lipid nanoparticle delivery was demonstrated in vivo. These highly efficient delivery strategies resulted in high levels of activation in multiple tissues. Finally, we demonstrate durable gene activation in vivo via transient delivery of a single dose of a gene activator that combines VP64, p65, and HSF1 with a SWI/SNF chromatin remodeling complex component SS18, representing an important step toward gene-activation-based therapeutics. This induced sustained gene activation could be inhibited via mRNA-encoded AcrIIA4, further improving the safety profile of this approach.
Authors
Beyersdorf, JP; Bawage, S; Iglesias, N; Peck, HE; Hobbs, RA; Wroe, JA; Zurla, C; Gersbach, CA; Santangelo, PJ
MLA Citation
Beyersdorf, Jared P., et al. “Robust, Durable Gene Activation In Vivo via mRNA-Encoded Activators.Acs Nano, Mar. 2022. Epmc, doi:10.1021/acsnano.1c10631.
URI
https://scholars.duke.edu/individual/pub1515882
PMID
35357116
Source
epmc
Published In
Acs Nano
Published Date
DOI
10.1021/acsnano.1c10631

Cas9-specific immune responses compromise local and systemic AAV CRISPR therapy in multiple dystrophic canine models.

Adeno-associated virus (AAV)-mediated CRISPR-Cas9 editing holds promise to treat many diseases. The immune response to bacterial-derived Cas9 has been speculated as a hurdle for AAV-CRISPR therapy. However, immunological consequences of AAV-mediated Cas9 expression have thus far not been thoroughly investigated in large mammals. We evaluate Cas9-specific immune responses in canine models of Duchenne muscular dystrophy (DMD) following intramuscular and intravenous AAV-CRISPR therapy. Treatment results initially in robust dystrophin restoration in affected dogs but also induces muscle inflammation, and Cas9-specific humoral and cytotoxic T-lymphocyte (CTL) responses that are not prevented by the muscle-specific promoter and transient prednisolone immune suppression. In normal dogs, AAV-mediated Cas9 expression induces similar, though milder, immune responses. In contrast, other therapeutic (micro-dystrophin and SERCA2a) and reporter (alkaline phosphatase, AP) vectors result in persistent expression without inducing muscle inflammation. Our results suggest Cas9 immunity may represent a critical barrier for AAV-CRISPR therapy in large mammals.
Authors
Hakim, CH; Kumar, SRP; Pérez-López, DO; Wasala, NB; Zhang, D; Yue, Y; Teixeira, J; Pan, X; Zhang, K; Million, ED; Nelson, CE; Metzger, S; Han, J; Louderman, JA; Schmidt, F; Feng, F; Grimm, D; Smith, BF; Yao, G; Yang, NN; Gersbach, CA; Chen, S-J; Herzog, RW; Duan, D
MLA Citation
Hakim, Chady H., et al. “Cas9-specific immune responses compromise local and systemic AAV CRISPR therapy in multiple dystrophic canine models.Nature Communications, vol. 12, no. 1, Nov. 2021, p. 6769. Epmc, doi:10.1038/s41467-021-26830-7.
URI
https://scholars.duke.edu/individual/pub1502787
PMID
34819506
Source
epmc
Published In
Nature Communications
Volume
12
Published Date
Start Page
6769
DOI
10.1038/s41467-021-26830-7

Gene delivery into cells and tissues

There are significant engineering challenges in translating gene and nucleic acid delivery from cell and animal models into the clinic. Off-target effects and inefficient delivery to the proper intracellular compartment of the targeted cells are major obstacles to success. Systemic delivery of any viral or nonviral vector requires an appreciation of the adverse physiological barriers that exist in vivo and incorporation of well-designed vector components that limit nonspecific uptake and accelerated clearance. In addition, the genetic engineer should consider the design criteria related to cell-specific action. For example, cell-surface recognition can increase the therapeutic index of a DNA- or RNA-based medication. Finally, one must consider the mechanism of cellular internalization, and investigators should attempt to target pathways and incorporate vector functionalities that will mediate trafficking to the subcellular compartment that is optimal for activity of the nucleic acid cargo. This chapter discusses key aspects of biodistribution and cellular uptake of nanoparticulate vectors and vehicles. It also surveys current strategies for engineering effective viral and nonviral packaging systems, systems for both targeted, systemic delivery and controlled, local release of nucleic acids or genes from engineered scaffolds, and strategies for directing the intracellular trafficking of vehicle contents to the nucleus or cytoplasm of target cells. The chapter concludes with the current clinical state of gene delivery for tissue engineering.
Authors
Nelson, CE; Duvall, CL; Prokop, A; Gersbach, CA; Davidson, JM
MLA Citation
Nelson, C. E., et al. “Gene delivery into cells and tissues.” Principles of Tissue Engineering, 2020, pp. 519–54. Scopus, doi:10.1016/B978-0-12-818422-6.00030-7.
URI
https://scholars.duke.edu/individual/pub1511024
Source
scopus
Published Date
Start Page
519
End Page
554
DOI
10.1016/B978-0-12-818422-6.00030-7

Full-length dystrophin restoration via targeted exon integration by AAV-CRISPR in a humanized mouse model of Duchenne muscular dystrophy.

Targeted gene-editing strategies have emerged as promising therapeutic approaches for the permanent treatment of inherited genetic diseases. However, precise gene correction and insertion approaches using homology-directed repair are still limited by low efficiencies. Consequently, many gene-editing strategies have focused on removal or disruption, rather than repair, of genomic DNA. In contrast, homology-independent targeted integration (HITI) has been reported to effectively insert DNA sequences at targeted genomic loci. This approach could be particularly useful for restoring full-length sequences of genes affected by a spectrum of mutations that are also too large to deliver by conventional adeno-associated virus (AAV) vectors. Here, we utilize an AAV-based, HITI-mediated approach for correction of full-length dystrophin expression in a humanized mouse model of Duchenne muscular dystrophy (DMD). We co-deliver CRISPR-Cas9 and a donor DNA sequence to insert the missing human exon 52 into its corresponding position within the DMD gene and achieve full-length dystrophin correction in skeletal and cardiac muscle. Additionally, as a proof-of-concept strategy to correct genetic mutations characterized by diverse patient mutations, we deliver a superexon donor encoding the last 28 exons of the DMD gene as a therapeutic strategy to restore full-length dystrophin in >20% of the DMD patient population. This work highlights the potential of HITI-mediated gene correction for diverse DMD mutations and advances genome editing toward realizing the promise of full-length gene restoration to treat genetic disease.
Authors
Pickar-Oliver, A; Gough, V; Bohning, JD; Liu, S; Robinson-Hamm, JN; Daniels, H; Majoros, WH; Devlin, G; Asokan, A; Gersbach, CA
MLA Citation
Pickar-Oliver, Adrian, et al. “Full-length dystrophin restoration via targeted exon integration by AAV-CRISPR in a humanized mouse model of Duchenne muscular dystrophy.Mol Ther, vol. 29, no. 11, Nov. 2021, pp. 3243–57. Pubmed, doi:10.1016/j.ymthe.2021.09.003.
URI
https://scholars.duke.edu/individual/pub1497106
PMID
34509668
Source
pubmed
Published In
Molecular Therapy : the Journal of the American Society of Gene Therapy
Volume
29
Published Date
Start Page
3243
End Page
3257
DOI
10.1016/j.ymthe.2021.09.003

Chromatin Remodeling of Colorectal Cancer Liver Metastasis is Mediated by an HGF-PU.1-DPP4 Axis.

Colorectal cancer (CRC) metastasizes mainly to the liver, which accounts for the majority of CRC-related deaths. Here it is shown that metastatic cells undergo specific chromatin remodeling in the liver. Hepatic growth factor (HGF) induces phosphorylation of PU.1, a pioneer factor, which in turn binds and opens chromatin regions of downstream effector genes. PU.1 increases histone acetylation at the DPP4 locus. Precise epigenetic silencing by CRISPR/dCas9KRAB or CRISPR/dCas9HDAC revealed that individual PU.1-remodeled regulatory elements collectively modulate DPP4 expression and liver metastasis growth. Genetic silencing or pharmacological inhibition of each factor along this chromatin remodeling axis strongly suppressed liver metastasis. Therefore, microenvironment-induced epimutation is an important mechanism for metastatic tumor cells to grow in their new niche. This study presents a potential strategy to target chromatin remodeling in metastatic cancer and the promise of repurposing drugs to treat metastasis.
Authors
Wang, L; Wang, E; Prado Balcazar, J; Wu, Z; Xiang, K; Wang, Y; Huang, Q; Negrete, M; Chen, K-Y; Li, W; Fu, Y; Dohlman, A; Mines, R; Zhang, L; Kobayashi, Y; Chen, T; Shi, G; Shen, JP; Kopetz, S; Tata, PR; Moreno, V; Gersbach, C; Crawford, G; Hsu, D; Huang, E; Bu, P; Shen, X
MLA Citation
Wang, Lihua, et al. “Chromatin Remodeling of Colorectal Cancer Liver Metastasis is Mediated by an HGF-PU.1-DPP4 Axis.Adv Sci (Weinh), vol. 8, no. 19, Oct. 2021, p. e2004673. Pubmed, doi:10.1002/advs.202004673.
URI
https://scholars.duke.edu/individual/pub1493278
PMID
34378358
Source
pubmed
Published In
Advanced Science (Weinheim, Baden Wurttemberg, Germany)
Volume
8
Published Date
Start Page
e2004673
DOI
10.1002/advs.202004673