593: Neonatal airway gene therapy delivery to enable effective adult dosing: A lentiviral vector study

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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Next Step in Gene Delivery: Modern Approaches and Further Perspectives of AAV Tropism Modification

Pharmaceutics ◽

10.3390/pharmaceutics13050750 ◽

2021 ◽

Vol 13 (5) ◽

pp. 750

Author(s):

Maxim A. Korneyenkov ◽

Andrey A. Zamyatnin

Keyword(s):

Gene Therapy ◽

Genome Organization ◽

Rational Design ◽

Structural Data ◽

Rational Approach ◽

Tissue Tropism ◽

Adeno Associated Virus ◽

Chemical Conjugation ◽

The Usa ◽

Therapy Delivery

Today, adeno-associated virus (AAV) is an extremely popular choice for gene therapy delivery. The safety profile and simplicity of the genome organization are the decisive advantages which allow us to claim that AAV is currently among the most promising vectors. Several drugs based on AAV have been approved in the USA and Europe, but AAV serotypes’ unspecific tissue tropism is still a serious limitation. In recent decades, several techniques have been developed to overcome this barrier, such as the rational design, directed evolution and chemical conjugation of targeting molecules with a capsid. Today, all of the abovementioned approaches confer the possibility to produce AAV capsids with tailored tropism, but recent data indicate that a better understanding of AAV biology and the growth of structural data may theoretically constitute a rational approach to most effectively produce highly selective and targeted AAV capsids. However, while we are still far from this goal, other approaches are still in play, despite their drawbacks and limitations.

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Intraoperative Use of Microscope-Integrated Optical Coherence Tomography for Subretinal Gene Therapy Delivery

Retina ◽

10.1097/iae.0000000000001646 ◽

2019 ◽

Vol 39 ◽

pp. S9-S12 ◽

Cited By ~ 15

Author(s):

Ninel Z. Gregori ◽

Byron L. Lam ◽

Janet L. Davis

Keyword(s):

Gene Therapy ◽

Optical Coherence Tomography ◽

Optical Coherence ◽

Integrated Optical ◽

Therapy Delivery

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904. CFTR Promoter in Lentiviral Vector System for Cystic Fibrosis Gene Therapy

Molecular Therapy ◽

10.1016/s1525-0016(16)40307-2 ◽

2008 ◽

Vol 16 ◽

pp. S337

Keyword(s):

Cystic Fibrosis ◽

Gene Therapy ◽

Lentiviral Vector ◽

Vector System ◽

Cystic Fibrosis Gene

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Combination gene therapy for HIV using a conditional suicidal gene with CCR5 knockout

Virology Journal ◽

10.1186/s12985-021-01501-7 ◽

2021 ◽

Vol 18 (1) ◽

Author(s):

Tugba Mehmetoglu-Gurbuz ◽

Rose Yeh ◽

Himanshu Garg ◽

Anjali Joshi

Keyword(s):

Gene Therapy ◽

Lentiviral Vector ◽

Suicide Gene ◽

Vector System ◽

Tat Protein ◽

Ccr5 Gene ◽

Hematopoietic Stem ◽

Therapy Strategy ◽

Hiv Tat ◽

A Cell

Abstract Background Gene therapy approaches using hematopoietic stem cells to generate an HIV resistant immune system have been shown to be successful. The deletion of HIV co-receptor CCR5 remains a viable strategy although co-receptor switching to CXCR4 remains a major pitfall. To overcome this, we designed a dual gene therapy strategy that incorporates a conditional suicide gene and CCR5 knockout (KO) to overcome the limitations of CCR5 KO alone. Methods A two-vector system was designed that included an integrating lentiviral vector that expresses a HIV Tat dependent Thymidine Kinase mutant SR39 (TK-SR39) and GFP reporter gene. The second non-integrating lentiviral (NIL) vector expresses a CCR5gRNA-CRISPR/Cas9 cassette and HIV Tat protein. Results Transduction of cells sequentially with the integrating followed by the NIL vector allows for insertion of the conditional suicide gene, KO of CCR5 and transient expression of GFP to enrich the modified cells. We used this strategy to modify TZM cells and generate a cell line that was resistant to CCR5 tropic viruses while permitting infection of CXCR4 tropic viruses which could be controlled via treatment with Ganciclovir. Conclusions Our study demonstrates proof of principle that a combination gene therapy for HIV is a viable strategy and can overcome the limitation of editing CCR5 gene alone.

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A self-inactivating lentiviral vector for SCID-X1 gene therapy that does not activate LMO2 expression in human T cells

Blood ◽

10.1182/blood-2009-10-250209 ◽

2010 ◽

Vol 116 (6) ◽

pp. 900-908 ◽

Cited By ~ 89

Author(s):

Sheng Zhou ◽

Disha Mody ◽

Suk See DeRavin ◽

Julia Hauer ◽

Taihe Lu ◽

...

Keyword(s):

Gene Therapy ◽

Bone Marrow ◽

Protein Expression ◽

Genomic Sequence ◽

Bone Marrow Cells ◽

Lentiviral Vector ◽

Severe Combined Immunodeficiency ◽

Vitro Assay ◽

Activation Assay ◽

Marrow Cells

Abstract To develop safer and more effective vectors for gene therapy of X-linked severe combined immunodeficiency (SCID-X1), we have evaluated new self-inactivating lentiviral vectors based on the HIV virus. The CL20i4-hγc-Revgen vector contains the entire human common γ chain (γc) genomic sequence driven by the γc promoter. The CL20i4-EF1α-hγcOPT vector uses a promoter fragment from the eukaryotic elongation factor alpha (EF1α) gene to express a codon-optimized human γc cDNA. Both vectors contain a 400-bp insulator fragment from the chicken β-globin locus within the self-inactivating long-terminal repeat. Transduction of bone marrow cells using either of these vectors restored T, B, and natural killer lymphocyte development and function in a mouse SCID-X1 transplantation model. Transduction of human CD34+ bone marrow cells from SCID-X1 patients with either vector restored T-cell development in an in vitro assay. In safety studies using a Jurkat LMO2 activation assay, only the CL20i4-EF1α-hγcOPT vector lacked the ability to transactivate LMO2 protein expression, whereas the CL20i4-hγc-Revgen vector significantly activated LMO2 protein expression. In addition, the CL20i4-EF1α-hγcOPT vector has not caused any tumors in transplanted mice. We conclude that the CL20i4-EF1α-hγcOPT vector may be suitable for testing in a clinical trial based on these preclinical demonstrations of efficacy and safety.

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