Patient-Customized Oligonucleotide Therapy for a Rare Genetic Disease

Kim, Jin‐Kuk; Hu, Chunguang; Achkar, Christelle Moufawad El; Black, Lauren E.; Douville, Julie; Larson, Austin; Pendergast, Mary K.; Goldkind, Sara F.; Lee, Eunjung A.; Kuniholm, Ashley; Soucy, Aubrie; Vaze, Jai; Belur, Nandkishore R.; Fredriksen, Kristina; Stojkovska, Iva; Tsytsykova, Alla V.; Armant, Myriam; DiDonato, Renata L.; Choi, Jae Young; Cornelissen, Laura; Pereira, Luis M.; Augustine, Erika F.; Genetti, Casie A.; Dies, Kira; Barton, Brenda; Williams, Lucinda; Goodlett, Benjamin D.; Riley, Bobbie; Pasternak, Amy; Berry, Emily R.; Pflock, Kelly A.; Chu, Stephen J; Reed, Chantal; Tyndall, Kimberly; Agrawal, Pankaj B.; Beggs, Alan H.; Grant, P. Ellen; Urion, David K.; Snyder, Richard O.; Waisbren, Susan E.; Poduri, Annapurna; Park, Peter J.; Patterson, Al; Biffi, Alessandra; Mazzulli, Joseph R.; Bodamer, Olaf A.; Berde, Charles B.; Yu, Timothy W.

doi:10.1056/nejmoa1813279

Cited by 576 publications

(461 citation statements)

References 25 publications

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“…To prevent the susceptibility and immunostimulatory activity of unmodified nucleic acids in vivo, we sought to investigate the tolerability of Cas proteins to chemically-stabilized DNA molecules. We introduced native and FDA-approved phosphorothioate-modified DNA molecules to the enzyme CRISPR-Cas12a [26][27][28] . The Cas12a from Lachnospiraceae bacterium ND2006 (LbaCas12a) assembled with guide CRISPR RNA sequences (crRNAs) recognizes 1) a T nucleotide-rich protospacer-adjacent motif (PAM) to target double-stranded DNA (dsDNA), or 2) single-stranded DNA (ssDNA) through sequence complementarity in a PAM-independent manner, and unleashes a robust, nonspecific ssDNA trans-cleavage activity that can be monitored using a fluorophore (F)-quencher (Q)-labeled reporter 17 ( Fig.…”

Section: Resultsmentioning

confidence: 99%

CRISPR-Cas-amplified urine biomarkers for multiplexed and portable cancer diagnostics

Hao

Zhao

Ngambenjawong

et al. 2020

Preprint

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Synthetic biomarkers, exogenous probes that generate molecular reporters, represent an emerging paradigm in precision diagnostics with applications across infectious and noncommunicable diseases. In order to achieve their promise, these methods reply on multiplexing strategies to provide tools that are both sensitive and specific. However, the field of synthetic biomarkers has not benefited from molecular strategies such as DNA-barcoding due to the susceptibility of nucleic acids in vivo. Herein, we exploit chemically-stabilized DNAs to tag synthetic biomarkers and produce diagnostic signals via CRISPR nucleases. Our strategy capitalizes on disease-associated, protease-activated release of nucleic acid barcodes and polymerase-amplification-free, CRISPR-Cas-mediated barcode detection in unprocessed biofluids. In murine cancer models, we show that the DNA-encoded urine biomarkers can noninvasively detect and monitor disease progression, and demonstrate that nuclease amplification can be harnessed to convert the readout to a point-of-care tool. This technique combines specificity with ease of use to offer a new platform to study human disease and guide therapeutic decisions.

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Section: Resultsmentioning

confidence: 99%

CRISPR-Cas-amplified urine biomarkers for multiplexed and portable cancer diagnostics

Hao

Zhao

Ngambenjawong

et al. 2020

Preprint

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“…Furthermore, each ASO that targets a particular gene is different so each one needs to be carefully assessed in vitro and in vivo to understand its half‐life and knockdown efficiency, and tissue distribution after intrathecal infusion. Importantly, several pioneering trials, most notably in Huntington's disease, where ASO targeting mRNA from HTT have been optimized (dose, concentration, time, delivery) in primates and humans, and can form a template for other disorders, including that for only a single patient, one example of which recently had minor but significant improvements in a child with an NDD.…”

Section: Drug Screening In Ipsc‐derived Cells For Translational Medicinementioning

confidence: 99%

A roadmap for neurodevelopmental disease modeling for non-stem cell biologists

Ernst

2020

Stem Cells Translational Medicine

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Stem and derivative cells induced from somatic tissues are a critical tool for disease modeling but significant technical hurdles hamper their use. The purpose of this review is to provide an overview of pitfalls and mitigation strategies for the nonstem cell biologist using induced pluripotent stem cells and investigating neurodevelopmental disorders. What sample sizes are reasonable? What derivation and purification protocols should be used to make human neurons? In what way should gene editing technologies be used to support discoveries? What kinds of preclinical studies are the most feasible? It is hoped that this roadmap will provide the necessary details for experimental planning and execution for those less familiar in the area of stem cell disease modeling. High-quality human preclinical models will allow for the discovery of molecular and cellular phenotypes specific to different neurodevelopmental disorders, and may provide the assays to advance translational medicine for unmet medical needs. K E Y W O R D S genetic engineering, neurodevelopmental disorders, neuronal induction, somatic cell reprogramming, stem cells, therapeutics

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“…However, recent developments suggest that there is now a clearer path to success . The high degree of personalization of therapy using this approach is illustrated in a recent report of the management of a rare genetic neurodegenerative disease, from diagnosis to synthesis and evaluation of a new oligonucleotide therapy in a single‐patient (N of 1) trial …”

Section: Commentmentioning

confidence: 99%

“…Readers of recent issues of the New England Journal of Medicine can be forgiven for having the same thoughts about genomic medicine. In one of the recent issues, for example, four of the five main articles had molecular genetics at their core, and two accompanying editorials commented on their implications, raising the thorny issues of regulatory approval, cost‐effectiveness and timely clinical adoption . A further article discussed the modelling of the placenta with stem cells, the new marvellous hydra.…”

Section: Commentmentioning

confidence: 99%

Genomic research delivering on promises: From rejuvenation to vaccines and pharmacogenetics

2020

J Clin Pharm Ther

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What is known and Objective There has been astounding progress made in the treatment of disease over recent years. This progress is particularly marked in cell therapy and in the personalization of therapy based on genetic insight, an approach known as genomic medicine. Our objective is to comment on the progress made in cell and genomic medicine against an historical backcloth of the search for rejuvenation. Comment In 1741, close to seven decades after Antoine van Leeuwenhoek first saw his microscopic animalcules, Abraham Trembley, a tutor in Leiden, reported on an organism that could regenerate itself. The strange organism was thought to hold the secret of life. If it does, we have yet to prise the secret out. However, the ensuing study of cell programming and induced stem cells has shed considerable light on cellular development and provided new insights on the rejuvenative capacity of organisms. Inventive scientists have provided a deeper understanding of cell replication and, from this, developed new medicines for an increasing range of diseases. Targeted therapies, oligonucleotide therapy, therapeutic monoclonal antibodies and pharmacogenetics are all new therapeutic areas originating from the improved insights. More will surely follow. What is new and conclusion Immortality is for the gods, but man's search for its elusive secrets, perhaps as old as man himself, will continue. Huge leaps have been made, and effective medicines have been developed from our improved insights into the mechanism of life. However, only the foolish will predict how far this new knowledge will lead us, and more particularly, at what speed new therapies will follow.

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Patient-Customized Oligonucleotide Therapy for a Rare Genetic Disease

Cited by 576 publications

References 25 publications

CRISPR-Cas-amplified urine biomarkers for multiplexed and portable cancer diagnostics

CRISPR-Cas-amplified urine biomarkers for multiplexed and portable cancer diagnostics

A roadmap for neurodevelopmental disease modeling for non-stem cell biologists

Genomic research delivering on promises: From rejuvenation to vaccines and pharmacogenetics

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