61. Successful Gene Therapy of a Mouse Model of Congenital Erythropoietic Porphyria with an Erythroid-Specific Lentiviral Vector

AbstractAmyotrophic lateral sclerosis (ALS) is an intractable neurodegenerative disease. CD68-positive bone marrow (BM)-derived cells (BMDCs) accumulate in the pathological lesion in the SOD1(G93A) ALS mouse model after BM transplantation (BMT). Therefore, we investigated whether BMDCs can be applied as gene carriers for cell-based gene therapy by employing the accumulation of BMDCs. In ALS mice, YFP reporter signals were observed in 12–14% of white blood cells (WBCs) and in the spinal cord via transplantation of BM after lentiviral vector (LV) infection. After confirmation of gene transduction by LV with the CD68 promoter in 4–7% of WBCs and in the spinal cord of ALS mice, BM cells were infected with LVs expressing glutamate transporter (GLT) 1 that protects neurons from glutamate toxicity, driven by the CD68 promoter, which were transplanted into ALS mice. The treated mice showed improvement of motor behaviors and prolonged survival. Additionally, interleukin (IL)-1β was significantly suppressed, and IL-4, arginase 1, and FIZZ were significantly increased in the mice. These results suggested that GLT1 expression by BMDCs improved the spinal cord environment. Therefore, our gene therapy strategy may be applied to treat neurodegenerative diseases such as ALS in which BMDCs accumulate in the pathological lesion by BMT.

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Faculty Opinions recommendation of Successful gene therapy of Diamond-Blackfan anemia in a mouse model and human CD34+ cord blood hematopoietic stem cells using a clinically applicable lentiviral vector.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.739368425.793582178 ◽

2021 ◽

Author(s):

Frank Staal

Keyword(s):

Stem Cells ◽

Gene Therapy ◽

Mouse Model ◽

Hematopoietic Stem Cells ◽

Cord Blood ◽

Lentiviral Vector ◽

Hematopoietic Stem ◽

Diamond Blackfan Anemia ◽

Human Cd34

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636. Lentiviral Vector-Mediated Gene Therapy for a Mouse Model of Spinal Muscular Atrophy

Molecular Therapy ◽

10.1016/s1525-0016(16)41078-6 ◽

2003 ◽

Vol 7 (5) ◽

pp. S248

Keyword(s):

Gene Therapy ◽

Mouse Model ◽

Spinal Muscular Atrophy ◽

Lentiviral Vector ◽

Muscular Atrophy

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Publisher Correction: In Utero Gene Therapy (IUGT) Using GLOBE Lentiviral Vector Phenotypically Corrects the Heterozygous Humanised Mouse Model and Its Progress Can Be Monitored Using MRI Techniques

Scientific Reports ◽

10.1038/s41598-019-55754-y ◽

2019 ◽

Vol 9 (1) ◽

Author(s):

Panicos Shangaris ◽

Stavros P. Loukogeorgakis ◽

Sindhu Subramaniam ◽

Christina Flouri ◽

Laurence H. Jackson ◽

...

Keyword(s):

Gene Therapy ◽

Mouse Model ◽

Lentiviral Vector ◽

In Utero

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

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Gene Therapy with pCCLchimP47 -phox Lentiviral Vector Achieves Long-term Reconstitution of NADPH Oxidase Activity in p47 -phox Chronic Granulomatous Disease (CGD) Mouse Model

Journal of Allergy and Clinical Immunology ◽

10.1016/j.jaci.2017.12.623 ◽

2018 ◽

Vol 141 (2) ◽

pp. AB197

Author(s):

Walmir Cutrim Aragao Filho ◽

Andrea Shejtman ◽

Marta Zinicola ◽

Ulrich Siler ◽

Janine Reichenbach ◽

...

Keyword(s):

Gene Therapy ◽

Mouse Model ◽

Nadph Oxidase ◽

Chronic Granulomatous Disease ◽

Oxidase Activity ◽

Lentiviral Vector ◽

Granulomatous Disease ◽

Nadph Oxidase Activity

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Hematopoietic Tumors in a Mouse Model of X-linked Chronic Granulomatous Disease after Lentiviral Vector-Mediated Gene Therapy

Molecular Therapy ◽

10.1016/j.ymthe.2020.09.030 ◽

2020 ◽

Author(s):

Raisa Jofra Hernández ◽

Andrea Calabria ◽

Francesca Sanvito ◽

Fabiola De Mattia ◽

Giada Farinelli ◽

...

Keyword(s):

Gene Therapy ◽

Mouse Model ◽

Chronic Granulomatous Disease ◽

Lentiviral Vector ◽

Granulomatous Disease

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Screening of Chimeric GAA Variants in a Preclinical Study of Pompe Disease Results in Candidate Vector for Hematopoietic Stem Cell Gene Therapy

10.1101/2021.12.28.474352 ◽

2021 ◽

Author(s):

Yildirim Dogan ◽

Cecilia N. Barese ◽

Jeffrey W. Schindler ◽

John K. Yoon ◽

Zeenath Unnisa ◽

...

Keyword(s):

Gene Therapy ◽

Mouse Model ◽

Pompe Disease ◽

Lentiviral Vector ◽

Therapeutic Potential ◽

Neuromuscular Disorder ◽

Current Standard ◽

Hematopoietic Stem ◽

Coding Sequences ◽

Enzyme Replacement

Pompe disease is a rare genetic neuromuscular disorder caused by acid alpha-glucosidase (GAA) deficiency resulting in lysosomal glycogen accumulation and progressive myopathy. Enzyme replacement therapy (ERT) is the current standard of care, which prolongs the quality of life for Pompe patients. However, ERT has limitations due to lack of enzyme penetration into the central nervous system (CNS) and skeletal muscles, immunogenicity against the recombinant enzyme, and requires life-long biweekly infusions. In a preclinical mouse model, a clinically relevant promoter to drive lentiviral vector-mediated expression of engineered GAA in autologous hematopoietic stem and progenitor cells (HSPC) was tested with nine unique human chimeric GAA coding sequences incorporating distinct peptide tags and codon-optimization iterations. Vectors including glycosylation independent lysosomal targeting (GILT) tags resulted in effective GAA enzyme delivery into key disease tissues with enhanced reduction of glycogen, myofiber and CNS vacuolation, compared to non-tagged GAA in Gaa knockout mice, a model of Pompe disease. Genetically modified microglial cells in brains were detected at low levels, but provided robust correction. Furthermore, an aminoacid substitution in the tag added to reduced capacity to induce insulin signaling and there was no evidence of off-target effects. This study demonstrated the therapeutic potential of lentiviral HSPC gene therapy exploiting optimized GAA tagged coding sequences to reverse Pompe disease pathology in a preclinical mouse model providing a promising vector candidate for further investigation.

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Successful gene therapy of Diamond-Blackfan anemia in a mouse model and human CD34+ cord blood hematopoietic stem cells using a clinically applicable lentiviral vector

Haematologica ◽

10.3324/haematol.2020.269142 ◽

2021 ◽

Author(s):

Yang Liu ◽

Maria Dahl ◽

Shubhranshu Debnath ◽

Michael Rothe ◽

Emma M. Smith ◽

...

Keyword(s):

Gene Therapy ◽

Bone Marrow ◽

Mouse Model ◽

Cord Blood ◽

Lentiviral Vector ◽

Bone Marrow Failure ◽

Hematopoietic Stem ◽

Diamond Blackfan Anemia ◽

Clinical Gene Therapy ◽

Cord Blood Cells

Diamond-Blackfan anemia (DBA) is an inherited bone marrow failure disorder with pure red blood cell aplasia associated with physical malformations and a predisposition to cancer. Twenty-five percent of patients with DBA have mutations in a gene encoding ribosomal protein S19 (RPS19). Our previous proof-of-concept studies demonstrated that DBA phenotype could be successfully treated using lentiviral vectors in Rps19-deficient DBA mice. In our present study, we developed a clinically applicable single gene self-inactivating lentiviral vector, containing the human RPS19 cDNA driven by the human elongation factor 1α short promoter, that can be used for clinical gene therapy development for RPS19-deficient DBA. We examined the efficacy and safety of the vector in a Rps19-deficient DBA mouse model and in human primary RPS19-deficient CD34+ cord blood cells. We observed that transduced Rps19-deficient bone marrow cells could reconstitute mice longterm and rescue the bone marrow failure and severe anemia observed in Rps19-deficient mice, with a low risk of mutagenesis and a highly polyclonal insertion site pattern. More importantly, the vector can also rescue impaired erythroid differentiation in human primary RPS19-deficient CD34+ cord blood hematopoietic stem cells. Collectively, our results demonstrate the efficacy and safety of using a clinically applicable lentiviral vector for the successful treatment of Rps19-deficient DBA in a mouse model and in human primary CD34+ cord blood cells. These findings show that this vector can be used to develop clinical gene therapy for RPS19-deficient DBA patients.

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