Challenges of CRISPR/Cas-Based Cell Therapy for Type 1 Diabetes: How Not to Engineer a “Trojan Horse”
Abstract
:1. Introduction
2. Etiology, Epidemiology, Diagnosis, and Treatment of Type 1 Diabetes
2.1. Etiology and Epidemiology of Type 1 Diabetes
2.2. Genetic Predisposition and Environment Triggers
2.3. T1D Diagnosis
2.4. T1D Treatment Options and Complications
2.5. T1D Cell Therapy
2.5.1. Sources for β-Cell Generation
2.5.2. HLA Haplotype Banks
2.6. Engineering of Immunoprivileged iPSC Lines
2.6.1. Safety Concerns and Approaches to Overcome Them
2.6.2. Challenges in CRISPR/Cas9-Based Generation of Immunoprivileged iPSCs
- 50 kb away from known gene, to prevent genes located nearby from being affected;
- 300 kb away from known oncogene, to prevent insertional oncogenesis;
- 300 kb away from miRNAs, 150 kb away from lncRNAs and tRNAs, so as not to disrupt gene expression and cell cycle regulation;
- 300 kb away from telomeres and centromeres, so as not to disrupt cell division;
- 20 kb away from known enhancer region, so as not to interfere with enhancer–gene interaction.
2.6.3. The Potential of Advanced CRISPR/Cas9 Tools for Engineering Immunoprivileged iPSC Lines
CRISPR/Cas9 Artificial Transcription Factors
Prime Editors
3. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Fasting Glucose Plasma (Fasting Is Defined as No Caloric Intake for at Least 8 h) | Random Plasma Glucose or 2-h Plasma Glucose during OGTT | Glycated Hemoglobin |
---|---|---|
≥7.0 mmol/L (126 mg/dL) | ≥11.1 mmol/L (200 mg/dL). | ≥6.5% (48 mmol/mol) |
Clinical Trial NCT | Company | Product | Cells | Administration |
---|---|---|---|---|
NCT03163511 | Viacyte | VC-02 | Pancreatic endoderm cells (PEC-01 cells) | Subcutaneous in a protective device |
NCT03513939 | Sernova | Cell Pouch | Therapeutic cells including islets | Abdominal musculature |
NCT04786262 | Vertex | VX-880 | Allogeneic fully differentiated insulin-producing islets | Infused into the hepatic portal vein |
NCT05565248 | CRISPR Therapeutics AG in collaboration with Viacyte | VCTX211 | Allogeneic pancreatic endoderm cells (PEC211) genetically modified using CRISPR/Cas9 | In a surgically implanted durable, removable, perforated device |
NCT05791201 | Vertex | VX-264 | Allogeneic fully differentiated insulin-producing islets | In a surgically implanted channel array protective device |
Type of Nuclease | Type of Delivery | Modification | Reference |
---|---|---|---|
spCas9n | Transfection plasmids | knockout B2M gene | [111] |
spCas9 | Nucleofection RNP | knockout CIITA gene | [113] |
knockout HLA-B, and CIITA genes | [125] | ||
knockout HLA-B, HLA-C, and CIITA genes | [124] | ||
knockout B2M, and CIITA genes | [123] | ||
knockout HLA-A, HLA-B, and HLA-DR | [168] | ||
Nucleofection plasmids | knockout HLA-B, HLA-C, and CIITA genes | [116] | |
knockout B2M, knockin HLA-G in B2M locus | [169] | ||
knockout HLA-B, HLA-C, and CIITA genes | [121] | ||
knockin PD-L1, HLA-G, CD47 in AAVS1 locus | |||
knockout CIITA gene | [122] | ||
knockout HLA-B gene | [170] | ||
knockout B2M gene | [171,172] | ||
knockout HLA-B, HLA-C, and CIITA genes | [116] | ||
knockin iC9 in AAVS1 | |||
knockout B2M, CIITA, and PVR genes | [173] | ||
Lipofection plasmids | knockout B2M gene | [112,174] | |
knockout B2M, and CIITA genes | [120] | ||
Lipofection gRNA in cells with knockin inducible Cas9 in AAVS1 locus | knockout HLA-A, HLA-B, HLA-C, and CIITA genes | [115] | |
Lentiviral particles | knockout B2M, and CIITA genes | [175] | |
Hi-Fi spCas9 | Nucleofection RNP | knockout HLA-A, HLA-B, and HLA-C | [176] |
knockout B2M gene | [128] | ||
Nucleofection plasmids | knockin HLA-E, PD-L1, IL-2 mutein, IL-10, and TGFB1 in GAPDH locus | [128] |
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Share and Cite
Karpov, D.S.; Sosnovtseva, A.O.; Pylina, S.V.; Bastrich, A.N.; Petrova, D.A.; Kovalev, M.A.; Shuvalova, A.I.; Eremkina, A.K.; Mokrysheva, N.G. Challenges of CRISPR/Cas-Based Cell Therapy for Type 1 Diabetes: How Not to Engineer a “Trojan Horse”. Int. J. Mol. Sci. 2023, 24, 17320. https://doi.org/10.3390/ijms242417320
Karpov DS, Sosnovtseva AO, Pylina SV, Bastrich AN, Petrova DA, Kovalev MA, Shuvalova AI, Eremkina AK, Mokrysheva NG. Challenges of CRISPR/Cas-Based Cell Therapy for Type 1 Diabetes: How Not to Engineer a “Trojan Horse”. International Journal of Molecular Sciences. 2023; 24(24):17320. https://doi.org/10.3390/ijms242417320
Chicago/Turabian StyleKarpov, Dmitry S., Anastasiia O. Sosnovtseva, Svetlana V. Pylina, Asya N. Bastrich, Darya A. Petrova, Maxim A. Kovalev, Anastasija I. Shuvalova, Anna K. Eremkina, and Natalia G. Mokrysheva. 2023. "Challenges of CRISPR/Cas-Based Cell Therapy for Type 1 Diabetes: How Not to Engineer a “Trojan Horse”" International Journal of Molecular Sciences 24, no. 24: 17320. https://doi.org/10.3390/ijms242417320