एक भ्रूण माउस मॉडल में एक उपन्यास Bioactive एजेंटों Intratracheal प्रशासन के लिए शल्य दृष्टिकोण
Summary
Bioactive एजेंटों के माउस भ्रूण में intratracheal प्रशासन के लिए एक उपन्यास शल्य दृष्टिकोण विकसित किया है. वितरण मार्ग आमतौर पर इस्तेमाल किया अंतर एमनियोटिक इंजेक्शन से भ्रूण माउस फेफड़ों को लक्षित करने में अधिक कुशल है. इस प्रक्रिया के लिए एक माउस मॉडल में नहीं वर्णित तारीख.
Abstract
Prenatal pulmonary delivery of cells, genes or pharmacologic agents could provide the basis for new therapeutic strategies for a variety of genetic and acquired diseases. Apart from congenital or inherited abnormalities with the requirement for long-term expression of the delivered gene, several non-inherited perinatal conditions, where short-term gene expression or pharmacological intervention is sufficient to achieve therapeutic effects, are considered as potential future indications for this kind of approach. Candidate diseases for the application of short-term prenatal therapy could be the transient neonatal deficiency of surfactant protein B causing neonatal respiratory distress syndrome1,2 or hyperoxic injuries of the neonatal lung3. Candidate diseases for permanent therapeutic correction are Cystic Fibrosis (CF)4, genetic variants of surfactant deficiencies5 and α1-antitrypsin deficiency6.
Generally, an important advantage of prenatal gene therapy is the ability to start therapeutic intervention early in development, at or even prior to clinical manifestations in the patient, thus preventing irreparable damage to the individual. In addition, fetal organs have an increased cell proliferation rate as compared to adult organs, which could allow a more efficient gene or stem cell transfer into the fetus. Furthermore, in utero gene delivery is performed when the individual’s immune system is not completely mature. Therefore, transplantation of heterologous cells or supplementation of a non-functional or absent protein with a correct version should not cause immune sensitization to the cell, vector or transgene product, which has recently been proven to be the case with both cellular and genetic therapies7.
In the present study, we investigated the potential to directly target the fetal trachea in a mouse model. This procedure is in use in larger animal models such as rabbits and sheep8, and even in a clinical setting9, but has to date not been performed before in a mouse model. When studying the potential of fetal gene therapy for genetic diseases such as CF, the mouse model is very useful as a first proof-of-concept because of the wide availability of different transgenic mouse strains, the well documented embryogenesis and fetal development, less stringent ethical regulations, short gestation and the large litter size.
Different access routes have been described to target the fetal rodent lung, including intra-amniotic injection10-12, (ultrasound-guided) intrapulmonary injection13,14 and intravenous administration into the yolk sac vessels15,16 or umbilical vein17. Our novel surgical procedure enables researchers to inject the agent of choice directly into the fetal mouse trachea which allows for a more efficient delivery to the airways than existing techniques18.
Protocol
Discussion
महत्वपूर्ण कदम
- हम साथ काम करने के लिए चुना गया माउस तनाव NMRI चूहों है क्योंकि वे पिल्ले (औसत कूड़े आकार 14.4 ± 1.8, स्वयं के डेटा) का एक प्रचुर संख्या है, हस्तक्षेप अच्छी तरह सहन और मातृ विशेषताओं अच्छा है. <…
Disclosures
The authors have nothing to disclose.
Acknowledgements
एम सी और AVdP डॉक्टरेट साथियों फ़्लैंडर्स में विज्ञान और प्रौद्योगिकी (आईडब्ल्यूटी Vlaanderen) के माध्यम से की अभिनव संवर्धन संस्थान से अनुदान द्वारा समर्थित हैं. संयुक्त अंशकालिक UZ लोवेन से क्लीनिकल रिसर्च फैलोशिप (KOOR) रखती है. DV एक डॉक्टरेट साथी यू लोवेन, DBOF/10/062 से एक अनुदान द्वारा समर्थित है. MMdC एक डॉक्टरेट साथी Conselho Nacional de Pesquisa ई (CNPq) Desenvolvimento और इरास्मस Mundus से एक अनुदान द्वारा समर्थित है. आईडब्ल्यूटी-Vlaanderen अनुसंधान द्वारा वित्त पोषित किया गया था, चुनाव आयोग अनुदान DIMI (LSHB सीटी 2005 ५,१२,१४६) और vivo आण्विक यू लोवेन से इमेजिंग अनुसंधान समूह (IMIR) द्वारा. हम UPenn वेक्टर उनके AAV6.2 प्लाज्मिड rAAV वेक्टर उत्पादन के लिए पैकेजिंग की तरह उपहार के लिए जेम्स एम. विल्सन द्वारा स्थापित कोर को स्वीकार करना चाहते हैं.
Materials
| Name of the reagent | Company | Catalogue number | Comments (optional) |
| NMRI mice | Janvier, Le Genest St Isle, France | ||
| Isoflurane | Isoba, Intervet / Schering-Plough Animal Health, Milton Keynes, UK | ||
| Prolene 6-0 | Ethicon, Groot Bijgaarden, Belgium | ||
| Vicryl 5-0 | Ethicon, Groot Bijgaarden, Belgium | ||
| 50 μl Hamilton Glass Syringe, Model 1710.5 TLLX SYR | Hamilton, Reno, NV, USA | 5495-20 | |
| 30G sharp needle | Hamilton, Reno, NV, USA | 7762-03 | |
| 2% xylocaine | AstraZeneca, Zoetermeer, The Netherlands |
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