Genetic Modification of Airway Progenitors after Lentiviral Gene Delivery to the Amniotic Fluid of Murine Fetuses

Mishra, Suparna; Wang, Xingchao; Smiley, Nancy; Xia, Ping; Hong, Chang; Senadheera, Dinithi; Bui, Kevin; Lutzko, Carolyn

doi:10.1165/rcmb.2009-0235oc

Cited by 8 publications

(10 citation statements)

References 40 publications

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“…Although our results should be interpreted with caution, due to small number of experimental animals, our data support the premise that in utero lung gene transfer using lung-targeting LVs can achieve high levels of pulmonary gene expression, which is maintained for up to 10 weeks. Life-long transduction of airway stem cells following intraamniotic delivery of LV has been successful in mice, 27 warranting longer-term studies (that is postnatal) in our sheep model using JSRVand VSVg-LV and minimally invasive methods of temporary fetal tracheal occlusion. Reversible balloon tracheal occlusion 49 is a potential strategy that could be applied to our fetal sheep lung gene transfer model to enable long-term postnatal studies.…”

Section: Discussionmentioning

confidence: 99%

“…In contrast, HIV-1-based lentiviral vectors (LVs) have increased packaging capacity (optimal transgene insert B5-7 Kb 23 ) and, following in utero delivery, are able to integrate into the host genome to provide life-long stable transgene expression. 16,[24][25][26][27] Fetal lung gene transfer has been investigated in non-human primates, 20,28 rabbits, 29 rats 30,31 and sheep 32 using vesicular stomatitis virus glycoprotein (VSVg) pseudotyped LV. Long-term luciferase transgene expression (that is 12-15 months after fetal gene transfer) has been reported 31,33 although there is a paucity of quantitative information on degree of epithelial transduction and cell types transduced.…”

Section: Introductionmentioning

confidence: 99%

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Jaagsiekte sheep retrovirus pseudotyped lentiviral vector-mediated gene transfer to fetal ovine lung

et al. 2011

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Viral vector-mediated gene transfer to the postnatal respiratory epithelium has, in general, been of low efficiency due to physical and immunological barriers, non-apical location of cellular receptors critical for viral uptake and limited transduction of resident stem/progenitor cells. These obstacles may be overcome using a prenatal strategy. In this study, HIV-1-based lentiviral vectors (LVs) pseudotyped with the envelope glycoproteins of Jaagsiekte sheep retrovirus (JSRV-LV), baculovirus GP64 (GP64-LV), Ebola Zaire-LV or vesicular stomatitis virus (VSVg-LV) and the adeno-associated virus-2/6.2 (AAV2/6.2) were compared for in utero transfer of a green fluorescent protein (GFP) reporter gene to ovine lung epithelium between days 65 and 78 of gestation. GFP expression was examined on day 85 or 136 of gestation (term is B145 days). The percentage of the respiratory epithelial cells expressing GFP in fetal sheep that received the JSRV-LV (3.18Â10 8 -6.85Â10 9 viral particles per fetus) was 24.6 ± 0.9% at 3 weeks postinjection (day 85) and 29.9±4.8% at 10 weeks postinjection (day 136). Expression was limited to the surface epithelium lining fetal airways o100 mm internal diameter. Fetal airways were amenable to VSVg-LV transduction, although the percentage of epithelial expression was low (6.6 ± 0.6%) at 1 week postinjection. GP64-LV, Ebola Zaire-LV and AAV2/6.2 failed to transduce the fetal ovine lung under these conditions. These data demonstrate that prenatal lung gene transfer with LV engineered to target apical surface receptors can provide sustained and high levels of transgene expression and support the therapeutic potential of prenatal gene transfer for the treatment of congenital lung diseases.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Jaagsiekte sheep retrovirus pseudotyped lentiviral vector-mediated gene transfer to fetal ovine lung

et al. 2011

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“…In contrast, after IA injections of VSV-G.HIV vector in fetal mice no lung transduction was observed (Buckley et al, 2008), while long-term (*1 year) airway transduction was demonstrated when using the baculovirus gp64-pseudotyped LV vector (Buckley et al, 2008), and in fetal rabbit lung transduction of tracheobronchial epithelium was seen within 4 days of delivery until 1 month (Skarsgard et al, 2005). Mishra and colleagues demonstrated that IAGT of VSV-G-pseudotyped LV vector on E14.5 or E16.5 resulted in long-term (7 months) transduction of epithelial cells of the small and large airways as well as transduction of long-lived respiratory epithelial progenitors with the ability to repair lung injury, suggesting a fetal development lung tropism for this vector (Mishra et al, 2011).…”

Section: Discussionmentioning

confidence: 99%

In Utero Lung Gene Transfer Using Adeno-Associated Viral and Lentiviral Vectors in Mice

Joyeux

Danzer²,

Limberis³

et al. 2014

Human Gene Therapy Methods

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Virus-mediated gene transfer to the fetal lung epithelium holds considerable promise for the therapeutic management of prenatally diagnosed, potentially life-threatening inherited lung diseases. In this study we hypothesized that efficient and life-long lung transduction can be achieved by in utero gene therapy, using viral vectors. To facilitate diffuse entry into the lung, viral vector was injected into the amniotic sac of C57BL/6 mice on embryonic day 16 (term, *20 days) in a volume of 10 ll. Vectors investigated included those based on adeno-associated virus (AAV) (serotypes 5, 6.2, 9, rh.64R1) and vesicular stomatitis virus G glycoprotein (VSV-G)-pseudotyped HIV-1-based lentivirus (LV). All vectors expressed green fluorescent protein (GFP) under the transcriptional control of various promoters including chicken b-actin (CB) or cytomegalovirus (CMV) for AAV and CMV or MND (myeloproliferative sarcoma virus enhancer, negative control region deleted) for LV. Pulmonary GFP gene expression was detected by fluorescence stereoscopic microscopy and immunohistochemistry for up to 9 months after birth. At equivalent vector doses (mean, 12 · 10 10 genome copies per fetus) three AAV vectors resulted in long-term (up to 9 months) pulmonary epithelium transduction. AAV2/6.2 transduced predominantly cells of the conducting airway epithelium, although transduction decreased 2 months after vector delivery. AAV2/9-transduced cells of the alveolar epithelium with a type 1 pneumocyte phenotype for up to 6 months. Although minimal levels of GFP expression were observed with AAV2/5 up to 9 months, the transduced cells immunostained positive for F480 and were retrievable by bronchoalveolar lavage, confirming an alveolar macrophage phenotype. No GFP expression was observed in lung epithelial cells after AAV2/rh.64R1 and VSV-G-LV vector-mediated gene transfer. We conclude that these experiments demonstrate that prenatal lung gene transfer with AAV vectors engineered to target pulmonary epithelial cells may provide sustained long-term levels of transgene expression, supporting the therapeutic potential of prenatal gene transfer for the treatment of congenital lung diseases.

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“…For patients with CF, this would mean ‘correcting’ the specific genetic mutation within the patients’ harvested stem cells prior to expansion, differentiation and seeding. As previously discussed, much research effort has been invested in developing methods to ‘correct’ the CFTR mutation (Conese et al ., ; Liu et al ., ; Mishra et al ., ) and these efforts will complement research in the fields of stem cell research and organ engineering.…”

Section: Tools and Approachesmentioning

confidence: 97%

Cell therapy for cystic fibrosis

Murphy

Atala

2013

J Tissue Eng Regen Med

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Currently there is no cure for cystic fibrosis (CF). Treatments are focused on addressing the disease symptoms, with varying degrees of success. Regenerative medicine holds the promise of regenerating dysfunctional or damaged tissues and to enhance the body's own endogenous repair mechanisms. The discovery of endogenous and exogenous stem cells has provided valuable tools for development of novel treatments for CF. The ability of stem cells to differentiate into functional pulmonary cells, modulate inflammatory responses and contribute to pulmonary function has provided researchers with multiple approaches to develop effective treatment strategies. Several approaches show promise to produce viable therapeutic treatments to treat the underlying cause of CF, reduce the symptoms and mitigate long-term damage, and generate functional replacement organs for end-stage transplantation. This review provides an overview of the rapidly progressing field of cell therapy for CF, focusing on the various cell types utilized and current strategies that show promise to improve life expectancy and quality of life for CF patients.

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Genetic Modification of Airway Progenitors after Lentiviral Gene Delivery to the Amniotic Fluid of Murine Fetuses

Cited by 8 publications

References 40 publications

Jaagsiekte sheep retrovirus pseudotyped lentiviral vector-mediated gene transfer to fetal ovine lung

Jaagsiekte sheep retrovirus pseudotyped lentiviral vector-mediated gene transfer to fetal ovine lung

In Utero Lung Gene Transfer Using Adeno-Associated Viral and Lentiviral Vectors in Mice

Cell therapy for cystic fibrosis

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