In Utero CRISPR Base Editing via Adenoviral Vector
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Solution Overview
Problem
Current genome editing technologies face challenges in achieving persistent high levels of edited cells postnatally due to immune responses and accessibility issues, particularly in treating genetic diseases with high prenatal or perinatal morbidity/mortality, where in utero editing offers a potential solution by leveraging the immunological immaturity of the fetus and high proliferative cells for efficient viral vector transduction.
Innovation Solution
Administration of an adenoviral vector carrying CRISPR-mediated base editor 3 (BE3) or base editor 4 (BE4) along with a guide RNA to a fetus for in utero genome editing, targeting specific mutations in therapeutic genes like Pcsk9 or Hpd, to introduce modified codons and treat genetic diseases before birth, thereby avoiding immune responses and enhancing editing efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If postnatal CRISPR-Cas9-mediated genome editing is performed, then disease-causing mutations can be corrected, but immune responses to viral vectors and bacterial Cas9 transgene product limit persistent high levels of edited cells
Solution Approach 1:
The patent performs genome editing in utero before the fetal immune system develops the capacity to mount responses against viral vectors and bacterial proteins. By administering the adenoviral vector carrying CRISPR-Cas9 and base editor components during fetal development, the editing occurs when immune tolerance is high, preventing subsequent immune-mediated clearance of edited cells that would occur with postnatal administration
Solution Approach 2:
The patent changes the temporal parameter of administration from postnatal to in utero, exploiting the developmental stage-dependent immune system characteristics. The fetal immune system's immaturity and tolerance to foreign antigens at this stage allows for efficient transduction and persistent expression without the harmful immune responses that would occur after birth
2Productivity
If postnatal genome editing is performed, then therapeutic correction can be achieved, but inaccessible and non-proliferative target cells reduce editing efficiency
Solution Approach 1:
The patent targets cells during fetal development when they are highly proliferative and accessible to viral vectors. By performing editing in utero, the treatment captures cells at a developmental stage when they are actively dividing and more susceptible to viral transduction, ensuring high editing efficiency before cells become inaccessible or non-proliferative in adult tissues
Solution Approach 2:
The patent exploits the dynamic state of fetal cells which are actively proliferating and metabolically active, making them highly receptive to viral vector delivery. The high turnover and accessibility of fetal target cells contrast with adult cells that may be quiescent or physically inaccessible, allowing for superior transduction and editing efficiency during the fetal period
3Quantity of substance
If high doses of viral vector are administered postnatally, then therapeutic dose can be achieved, but immune responses are triggered
Solution Approach 1:
The patent changes the administration timing parameter from postnatal to in utero, allowing delivery of high viral vector doses that would be immunogenic in adults. The fetal immune system's tolerance to foreign proteins and vectors enables administration of therapeutically effective high doses without triggering the immune clearance mechanisms that would eliminate both the vector and edited cells in postnatal subjects
Data Source
AI summary
A method for in utero genome editing, the method comprising administering to a subject an adenoviral vector, wherein the subject is a fetus, the adenoviral vector comprising CRISPR-mediated base editor and a guide RNA (gRNA), the gRNA targeting a mutation in a therapeutic gene; and introducing a modified codon in the therapeutic gene by base editing the therapeutic gene, wherein the base editing is performed by the adenoviral vector, an adeno-associated viral vector or lipid based nanoparticle.