In vivo engineered B cells secrete high titers of broadly neutralizing anti-HIV antibodies in mice.
Alessio David NahmadCicera R LazzarottoNatalie ZeliksonTalia KustinMary TenutaDeli HuangInbal ReuveniDaniel NatafYuval RavivMiriam Horovitz-FriedIris DotanYaron CarmiRina Rosin-ArbesfeldDavid NemazeeJames E VossAdi SternShengdar Q TsaiAdi BarzelPublished in: Nature biotechnology (2022)
Transplantation of B cells engineered ex vivo to secrete broadly neutralizing antibodies (bNAbs) has shown efficacy in disease models. However, clinical translation of this approach would require specialized medical centers, technically demanding protocols and major histocompatibility complex compatibility of donor cells and recipients. Here we report in vivo B cell engineering using two adeno-associated viral vectors, with one coding for Staphylococcus aureus Cas9 (saCas9) and the other for 3BNC117, an anti-HIV bNAb. After intravenously injecting the vectors into mice, we observe successful editing of B cells leading to memory retention and bNAb secretion at neutralizing titers of up to 6.8 µg ml -1 . We observed minimal clustered regularly interspaced palindromic repeats (CRISPR)-Cas9 off-target cleavage as detected by unbiased CHANGE-sequencing analysis, whereas on-target cleavage in undesired tissues is reduced by expressing saCas9 from a B cell-specific promoter. In vivo B cell engineering to express therapeutic antibodies is a safe, potent and scalable method, which may be applicable not only to infectious diseases but also in the treatment of noncommunicable conditions, such as cancer and autoimmune disease.
Keyphrases
- crispr cas
- genome editing
- infectious diseases
- antiretroviral therapy
- hiv positive
- hiv infected
- staphylococcus aureus
- dengue virus
- hiv testing
- human immunodeficiency virus
- hepatitis c virus
- hiv aids
- high fat diet induced
- gene therapy
- gene expression
- induced apoptosis
- men who have sex with men
- dna binding
- dna methylation
- papillary thyroid
- palliative care
- sars cov
- wild type
- single cell
- transcription factor
- multiple sclerosis
- cell cycle arrest
- type diabetes
- insulin resistance
- zika virus
- signaling pathway
- pseudomonas aeruginosa
- drug induced
- bone marrow
- cell proliferation
- squamous cell
- squamous cell carcinoma
- replacement therapy
- kidney transplantation
- methicillin resistant staphylococcus aureus