Immune-orthogonal orthologues of AAV capsids and of Cas9 circumvent the immune response to the administration of gene therapy

Moreno, Ana M.; Palmer, Nathan; Alemán, Fernando; Chen, Genghao; Pla, Andrew; Jiang, Ning; Chew, Wei Leong; Law, Mansun; Mali, Prashant

doi:10.1038/s41551-019-0431-2

Cited by 88 publications

(69 citation statements)

References 95 publications

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“…Thus the immune responses caused by these exogenous components are hard to evaluate accurately (Wei et al, 2016;Charlesworth et al, 2019). Possible solutions are to develop low immunogenic systems (Moreno et al, 2019) to have favorable biocompatibility by implanting encapsulated cells carrying synthetic circuits with biomaterials (Lathuiliere et al, 2014;Ye et al, 2017). Furthermore, as a genome-engineering tool, CRISPR-Cas9 system probably generates off-target effects at amplified loci (Munoz et al, 2016) and DNA damage is induced by Cas9.…”

Section: Discussionmentioning

confidence: 99%

Synthetic Biology Speeds Up Drug Target Discovery

Xie

Yang

et al. 2020

Front. Pharmacol.

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As a rising emerging field, synthetic biology intends to realize precise regulations of cellular network by constructing artificial synthetic circuits, and it brings great opportunities to treat diseases and discover novel drug targets. Depending on the combination mode of different logic gates, various synthetic circuits are created to carry out multilevel regulations. In given synthetic circuits, drugs often act as inputs to drive circuits operation. It is becoming available to construct drug-responsive gene circuits for experimentally treating various disease models, including metabolic disease, immunity disease, cancer and bacterial infection. Synthetic biology works well in association with the CRISPR system for drug target functional screening. Remarkably, more and more well-designed circuits are developed to discover novel drug targets and precisely regulate drug therapy for diseases.

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Section: Discussionmentioning

confidence: 99%

Synthetic Biology Speeds Up Drug Target Discovery

Xie

Yang

et al. 2020

Front. Pharmacol.

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“…For instance, CRISPR-Cas9 proteins have been a powerful tool for research in genetic and epigenetic engineering, but can evoke adaptive immune responses and tissue damage in vivo , and are therefore potentially pathogenic if applied to correct inherited genetic defects to treat diseases ( 32 ). Protein engineering to remove immunogenic epitopes and humanize these synthetic proteins to circumvent this issue can be difficult owing to the high diversity of the human leukocyte antigen (HLA) loci ( 33 ). Using our acoustogenetic approach, the transiently induced Hsp-driven synthetic regulators (e.g.…”

Section: Discussionmentioning

confidence: 99%

Acoustogenetic Control of CAR T Cells via Focused Ultrasound

Wu¹,

Huang³

et al. 2020

Preprint

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14Optogenetics can control specific molecular events in living systems, but the penetration depth of 15 light is typically limited at hundreds of micrometers. Focused ultrasound (FUS), on the other 16 hand, can deliver energy safely and noninvasively into tissues at depths of centimeters. Here we 17 have developed an acoustogenetic approach using short-pulsed FUS to remotely and directly 18 control the genetics and cellular functions of engineered mammalian cells for therapeutic 19 purposes. We applied this acoustogenetic approach to control chimeric antigen receptor (CAR) T 20 cells with high spatiotemporal precision, aiming to mitigate the potentially lethal "on-target off-21 tumor" effects of CAR T cell therapy. We first verified the controllability of our acoustogenetic 22 CAR T cells in recognizing and killing tumor cells in vitro, and then applied this approach in 23 vivo to suppress tumor growth of both lymphoma and prostate cancers. The results indicate that 24 FUS-based acoustogenetics can allow the noninvasive and remote activation, without any 25 exogenous cofactor, of different types of CAR T cells for cancer therapeutics. 26Optogenetics enables the control of specific molecular events and cellular functions in living 27 systems with high spatiotemporal resolutions. However, optogenetics cannot reach deep tissues, 28 with the penetration depth of light typically limited at micrometer to millimeter scales (1). 29Ultrasound can be focused to deliver mechanical energy safely and noninvasively into small 30 volumes of tissue deep inside the body up to tens of centimeters (1). The rapidly oscillating 31 pressure of focused ultrasound (FUS) waves and the resultant cycles of mechanical 32 loading/unloading can lead to local heat generation in biological tissues. Aided by Magnetic 33Resonance Imaging (MRI) thermometry, FUS has been widely applied to clinically ablate 34 tumors, and control drug delivery, vasodilation, neuromodulation (2), and transgene expression 35(3-5). Transcription factors and genetic circuits have also been engineered to convert the FUS-36 generated heat into genetic regulation to control microbial systems in vivo (6). However, there is 37 a lack of general methods using FUS to control mammalian cell functions in vivo for therapeutic 38 applications. 40Chimeric antigen receptor (CAR) T cell therapy, where T cells are genetically programmed with 41 redirected specificity against malignant cells, is becoming a paradigm-shifting approach for 42 cancer treatment, especially for blood cancers (7). However, major challenges remain for solid 43 tumors before CAR-based immunotherapy can be widely adopted. For instance, the non-specific 44 targeting of the CAR T cells against normal tissues (on-target off-tumor toxicities) can be life-45 threatening: off-tumor toxicities against the lung, the brain, and the heart have caused multiple 46 cases of deaths (7-10). Immunosuppressive corticosteroid therapy and suicide gene engineering 47 are relatively effective in suppressing off-tumor tox...

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“…To this end, several Cas9 orthologs and AAV serotypes were tested based on sequence similarities and predicted binding strength to MHC class I and class II to screen for immune orthologs that can be used for safe repeated administration of AAV-CRISPR gene therapy. Although no two AAV serotypes were found to completely circumvent immune recognition, the study verified 3 Cas9 orthologs [SpCas9, SaCas9, and Campylobacter jejuni Cas9 (CjCas9)] which showed robust editing efficiency and tolerated repeated administration due to reduced immunogenic toxicity in mice immunized against AAV and Cas9 (76). A major caveat is pre-existing immunity in humans against 2 of these orthologs-SpCas9 and SaCas9, leaving CjCas9 as the only current option for this cohort of patients.…”

Section: Immunotoxicitymentioning

confidence: 92%

CRISPR Gene Therapy: Applications, Limitations, and Implications for the Future

2020

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A series of recent discoveries harnessing the adaptive immune system of prokaryotes to perform targeted genome editing is having a transformative influence across the biological sciences. The discovery of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated (Cas) proteins has expanded the applications of genetic research in thousands of laboratories across the globe and is redefining our approach to gene therapy. Traditional gene therapy has raised some concerns, as its reliance on viral vector delivery of therapeutic transgenes can cause both insertional oncogenesis and immunogenic toxicity. While viral vectors remain a key delivery vehicle, CRISPR technology provides a relatively simple and efficient alternative for site-specific gene editing, obliviating some concerns raised by traditional gene therapy. Although it has apparent advantages, CRISPR/Cas9 brings its own set of limitations which must be addressed for safe and efficient clinical translation. This review focuses on the evolution of gene therapy and the role of CRISPR in shifting the gene therapy paradigm. We review the emerging data of recent gene therapy trials and consider the best strategy to move forward with this powerful but still relatively new technology.

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Immune-orthogonal orthologues of AAV capsids and of Cas9 circumvent the immune response to the administration of gene therapy

Cited by 88 publications

References 95 publications

Synthetic Biology Speeds Up Drug Target Discovery

Synthetic Biology Speeds Up Drug Target Discovery

Acoustogenetic Control of CAR T Cells via Focused Ultrasound

CRISPR Gene Therapy: Applications, Limitations, and Implications for the Future

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