Pulmonary Gene Silencing in Transgenic EGFP Mice Using Aerosolised Chitosan/siRNA Nanoparticles
Purpose This work describes the production and application of an aerosolised formulation of chitosan nanoparticles for improved pulmonary siRNA delivery and gene silencing in mice. Methods Aerosolised chitosan/siRNA nanoparticles were pneumatically formed using a nebulising catheter and sized by las...
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| Veröffentlicht in: | Pharmaceutical research 2010-12, Vol.27 (12), p.2520-2527 |
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| creator | Nielsen, Ebbe J. B Nielsen, Jan M Becker, Daniel Karlas, Alexander Prakash, Hridayesh Glud, Sys Z Merrison, Jonathan Besenbacher, Flemming Meyer, Thomas F Kjems, Jørgen Howard, Kenneth A |
| description | Purpose This work describes the production and application of an aerosolised formulation of chitosan nanoparticles for improved pulmonary siRNA delivery and gene silencing in mice. Methods Aerosolised chitosan/siRNA nanoparticles were pneumatically formed using a nebulising catheter and sized by laser diffraction. In vitro silencing of aerosolised and non-aerosolised formulations was evaluated in an EGFP endogenous-expressing H1299 cell line by flow cytometry. Non-invasive intratracheal insertion of the catheter was used to study nanoparticle deposition by histological detection of Cy3-labeled siRNA and gene silencing in transgenic EGFP mouse lungs using a flow cytometric method. Results Flow cytometric analysis demonstrated minimal alteration in gene silencing efficiency before (68%) and after (62%) aerosolisation in EGFP-expressing H1299 cells. Intratracheal catheter administration in mice resulted in nanoparticle deposition throughout the entire lung in both alveoli and bronchiolar regions using low amounts of siRNA. Transgenic EGFP mice dosed with the aerosolised nanoparticle formulation showed significant EGFP gene silencing (68% reduction compared to mismatch group). Conclusions This work provides a technology platform for effective pulmonary delivery and gene silencing of RNAi therapeutics with potential use in preclinical studies of respiratory disease treatment. |
| doi_str_mv | 10.1007/s11095-010-0255-y |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_883018778</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2193803661</sourcerecordid><originalsourceid>FETCH-LOGICAL-c492t-f55fbee2a8842bff0984c12942e313ce664b16ceb6b864b08599323984227ba83</originalsourceid><addsrcrecordid>eNqNkUFvEzEQhS1ERUPgB3CBFRdOS2ds7659jKI2IJW2ahuJm-V1Z4OrjTfY2UP-PY62gMQB1ZexNN97o5nH2DuEzwjQnCVE0FUJCCXwqioPL9gMq0aUGuT3l2wGDZelaiSestcpPQKAQi1fsVMOiksBesbMzdhvh2DjoVhRoOLO9xScD5vCh-I-2pA2FLwrzlcXN8U376hYp2N3QXFIQ-8TPRTLH34_JBvOkr-9WhRXNgw7G_fe9ZTesJPO9onePtU5W1-c3y-_lJfXq6_LxWXppOb7squqriXiVinJ264DraRDriUngcJRXcsWa0dt3ar8BVVpLbjIFOdNa5WYs0-T7y4OP0dKe7P1yVHf20DDmIxSAlA1zTNIULJGkd-cffyHfBzGGPIaGWqqWkslM4QT5PJBUqTO7KLf5nsaBHNMyUwpmZySOaZkDlnz_sl4bLf08EfxO5YM8AlIuRU2FP9O_p_rh0nU2cHYTfTJrO84YF5cI2qN4hfi06TY</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>807569484</pqid></control><display><type>article</type><title>Pulmonary Gene Silencing in Transgenic EGFP Mice Using Aerosolised Chitosan/siRNA Nanoparticles</title><source>MEDLINE</source><source>SpringerLink Journals - AutoHoldings</source><creator>Nielsen, Ebbe J. B ; Nielsen, Jan M ; Becker, Daniel ; Karlas, Alexander ; Prakash, Hridayesh ; Glud, Sys Z ; Merrison, Jonathan ; Besenbacher, Flemming ; Meyer, Thomas F ; Kjems, Jørgen ; Howard, Kenneth A</creator><creatorcontrib>Nielsen, Ebbe J. B ; Nielsen, Jan M ; Becker, Daniel ; Karlas, Alexander ; Prakash, Hridayesh ; Glud, Sys Z ; Merrison, Jonathan ; Besenbacher, Flemming ; Meyer, Thomas F ; Kjems, Jørgen ; Howard, Kenneth A</creatorcontrib><description>Purpose This work describes the production and application of an aerosolised formulation of chitosan nanoparticles for improved pulmonary siRNA delivery and gene silencing in mice. Methods Aerosolised chitosan/siRNA nanoparticles were pneumatically formed using a nebulising catheter and sized by laser diffraction. In vitro silencing of aerosolised and non-aerosolised formulations was evaluated in an EGFP endogenous-expressing H1299 cell line by flow cytometry. Non-invasive intratracheal insertion of the catheter was used to study nanoparticle deposition by histological detection of Cy3-labeled siRNA and gene silencing in transgenic EGFP mouse lungs using a flow cytometric method. Results Flow cytometric analysis demonstrated minimal alteration in gene silencing efficiency before (68%) and after (62%) aerosolisation in EGFP-expressing H1299 cells. Intratracheal catheter administration in mice resulted in nanoparticle deposition throughout the entire lung in both alveoli and bronchiolar regions using low amounts of siRNA. Transgenic EGFP mice dosed with the aerosolised nanoparticle formulation showed significant EGFP gene silencing (68% reduction compared to mismatch group). Conclusions This work provides a technology platform for effective pulmonary delivery and gene silencing of RNAi therapeutics with potential use in preclinical studies of respiratory disease treatment.</description><identifier>ISSN: 0724-8741</identifier><identifier>EISSN: 1573-904X</identifier><identifier>DOI: 10.1007/s11095-010-0255-y</identifier><identifier>PMID: 20824309</identifier><language>eng</language><publisher>Boston: Boston : Springer US</publisher><subject>aerosol ; Aerosols ; Alveoli ; Animals ; Base Sequence ; Biochemistry ; Biomedical and Life Sciences ; Biomedical Engineering and Bioengineering ; Biomedical materials ; Biomedicine ; Catheters ; Cell Line ; Chitosan ; Diffraction ; Flow Cytometry ; Gene Knockdown Techniques ; Gene Silencing ; Gene therapy ; Green Fluorescent Proteins - genetics ; Lasers ; Lung ; Medical Law ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Nanoparticles ; Pharmacology/Toxicology ; Pharmacy ; pulmonary gene silencing ; Research Paper ; Ribonucleic acid ; RNA ; RNA Interference ; RNA-mediated interference ; Rodents ; siRNA ; Trachea ; Transgenic animals ; Transgenic mice</subject><ispartof>Pharmaceutical research, 2010-12, Vol.27 (12), p.2520-2527</ispartof><rights>Springer Science+Business Media, LLC 2010</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c492t-f55fbee2a8842bff0984c12942e313ce664b16ceb6b864b08599323984227ba83</citedby><cites>FETCH-LOGICAL-c492t-f55fbee2a8842bff0984c12942e313ce664b16ceb6b864b08599323984227ba83</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s11095-010-0255-y$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s11095-010-0255-y$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27901,27902,41464,42533,51294</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20824309$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Nielsen, Ebbe J. B</creatorcontrib><creatorcontrib>Nielsen, Jan M</creatorcontrib><creatorcontrib>Becker, Daniel</creatorcontrib><creatorcontrib>Karlas, Alexander</creatorcontrib><creatorcontrib>Prakash, Hridayesh</creatorcontrib><creatorcontrib>Glud, Sys Z</creatorcontrib><creatorcontrib>Merrison, Jonathan</creatorcontrib><creatorcontrib>Besenbacher, Flemming</creatorcontrib><creatorcontrib>Meyer, Thomas F</creatorcontrib><creatorcontrib>Kjems, Jørgen</creatorcontrib><creatorcontrib>Howard, Kenneth A</creatorcontrib><title>Pulmonary Gene Silencing in Transgenic EGFP Mice Using Aerosolised Chitosan/siRNA Nanoparticles</title><title>Pharmaceutical research</title><addtitle>Pharm Res</addtitle><addtitle>Pharm Res</addtitle><description>Purpose This work describes the production and application of an aerosolised formulation of chitosan nanoparticles for improved pulmonary siRNA delivery and gene silencing in mice. Methods Aerosolised chitosan/siRNA nanoparticles were pneumatically formed using a nebulising catheter and sized by laser diffraction. In vitro silencing of aerosolised and non-aerosolised formulations was evaluated in an EGFP endogenous-expressing H1299 cell line by flow cytometry. Non-invasive intratracheal insertion of the catheter was used to study nanoparticle deposition by histological detection of Cy3-labeled siRNA and gene silencing in transgenic EGFP mouse lungs using a flow cytometric method. Results Flow cytometric analysis demonstrated minimal alteration in gene silencing efficiency before (68%) and after (62%) aerosolisation in EGFP-expressing H1299 cells. Intratracheal catheter administration in mice resulted in nanoparticle deposition throughout the entire lung in both alveoli and bronchiolar regions using low amounts of siRNA. Transgenic EGFP mice dosed with the aerosolised nanoparticle formulation showed significant EGFP gene silencing (68% reduction compared to mismatch group). Conclusions This work provides a technology platform for effective pulmonary delivery and gene silencing of RNAi therapeutics with potential use in preclinical studies of respiratory disease treatment.</description><subject>aerosol</subject><subject>Aerosols</subject><subject>Alveoli</subject><subject>Animals</subject><subject>Base Sequence</subject><subject>Biochemistry</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedical Engineering and Bioengineering</subject><subject>Biomedical materials</subject><subject>Biomedicine</subject><subject>Catheters</subject><subject>Cell Line</subject><subject>Chitosan</subject><subject>Diffraction</subject><subject>Flow Cytometry</subject><subject>Gene Knockdown Techniques</subject><subject>Gene Silencing</subject><subject>Gene therapy</subject><subject>Green Fluorescent Proteins - genetics</subject><subject>Lasers</subject><subject>Lung</subject><subject>Medical Law</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>Mice, Transgenic</subject><subject>Nanoparticles</subject><subject>Pharmacology/Toxicology</subject><subject>Pharmacy</subject><subject>pulmonary gene silencing</subject><subject>Research Paper</subject><subject>Ribonucleic acid</subject><subject>RNA</subject><subject>RNA Interference</subject><subject>RNA-mediated interference</subject><subject>Rodents</subject><subject>siRNA</subject><subject>Trachea</subject><subject>Transgenic animals</subject><subject>Transgenic mice</subject><issn>0724-8741</issn><issn>1573-904X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNqNkUFvEzEQhS1ERUPgB3CBFRdOS2ds7659jKI2IJW2ahuJm-V1Z4OrjTfY2UP-PY62gMQB1ZexNN97o5nH2DuEzwjQnCVE0FUJCCXwqioPL9gMq0aUGuT3l2wGDZelaiSestcpPQKAQi1fsVMOiksBesbMzdhvh2DjoVhRoOLO9xScD5vCh-I-2pA2FLwrzlcXN8U376hYp2N3QXFIQ-8TPRTLH34_JBvOkr-9WhRXNgw7G_fe9ZTesJPO9onePtU5W1-c3y-_lJfXq6_LxWXppOb7squqriXiVinJ264DraRDriUngcJRXcsWa0dt3ar8BVVpLbjIFOdNa5WYs0-T7y4OP0dKe7P1yVHf20DDmIxSAlA1zTNIULJGkd-cffyHfBzGGPIaGWqqWkslM4QT5PJBUqTO7KLf5nsaBHNMyUwpmZySOaZkDlnz_sl4bLf08EfxO5YM8AlIuRU2FP9O_p_rh0nU2cHYTfTJrO84YF5cI2qN4hfi06TY</recordid><startdate>20101201</startdate><enddate>20101201</enddate><creator>Nielsen, Ebbe J. B</creator><creator>Nielsen, Jan M</creator><creator>Becker, Daniel</creator><creator>Karlas, Alexander</creator><creator>Prakash, Hridayesh</creator><creator>Glud, Sys Z</creator><creator>Merrison, Jonathan</creator><creator>Besenbacher, Flemming</creator><creator>Meyer, Thomas F</creator><creator>Kjems, Jørgen</creator><creator>Howard, Kenneth A</creator><general>Boston : Springer US</general><general>Springer US</general><general>Springer Nature B.V</general><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7RV</scope><scope>7TK</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>BENPR</scope><scope>CCPQU</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>K9.</scope><scope>KB0</scope><scope>M0S</scope><scope>M1P</scope><scope>NAPCQ</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>7QO</scope><scope>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope></search><sort><creationdate>20101201</creationdate><title>Pulmonary Gene Silencing in Transgenic EGFP Mice Using Aerosolised Chitosan/siRNA Nanoparticles</title><author>Nielsen, Ebbe J. B ; Nielsen, Jan M ; Becker, Daniel ; Karlas, Alexander ; Prakash, Hridayesh ; Glud, Sys Z ; Merrison, Jonathan ; Besenbacher, Flemming ; Meyer, Thomas F ; Kjems, Jørgen ; Howard, Kenneth A</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c492t-f55fbee2a8842bff0984c12942e313ce664b16ceb6b864b08599323984227ba83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>aerosol</topic><topic>Aerosols</topic><topic>Alveoli</topic><topic>Animals</topic><topic>Base Sequence</topic><topic>Biochemistry</topic><topic>Biomedical and Life Sciences</topic><topic>Biomedical Engineering and Bioengineering</topic><topic>Biomedical materials</topic><topic>Biomedicine</topic><topic>Catheters</topic><topic>Cell Line</topic><topic>Chitosan</topic><topic>Diffraction</topic><topic>Flow Cytometry</topic><topic>Gene Knockdown Techniques</topic><topic>Gene Silencing</topic><topic>Gene therapy</topic><topic>Green Fluorescent Proteins - genetics</topic><topic>Lasers</topic><topic>Lung</topic><topic>Medical Law</topic><topic>Mice</topic><topic>Mice, Inbred C57BL</topic><topic>Mice, Transgenic</topic><topic>Nanoparticles</topic><topic>Pharmacology/Toxicology</topic><topic>Pharmacy</topic><topic>pulmonary gene silencing</topic><topic>Research Paper</topic><topic>Ribonucleic acid</topic><topic>RNA</topic><topic>RNA Interference</topic><topic>RNA-mediated interference</topic><topic>Rodents</topic><topic>siRNA</topic><topic>Trachea</topic><topic>Transgenic animals</topic><topic>Transgenic mice</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Nielsen, Ebbe J. B</creatorcontrib><creatorcontrib>Nielsen, Jan M</creatorcontrib><creatorcontrib>Becker, Daniel</creatorcontrib><creatorcontrib>Karlas, Alexander</creatorcontrib><creatorcontrib>Prakash, Hridayesh</creatorcontrib><creatorcontrib>Glud, Sys Z</creatorcontrib><creatorcontrib>Merrison, Jonathan</creatorcontrib><creatorcontrib>Besenbacher, Flemming</creatorcontrib><creatorcontrib>Meyer, Thomas F</creatorcontrib><creatorcontrib>Kjems, Jørgen</creatorcontrib><creatorcontrib>Howard, Kenneth A</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Nursing & Allied Health Database</collection><collection>Neurosciences Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central</collection><collection>ProQuest One Community College</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Nursing & Allied Health Premium</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>MEDLINE - Academic</collection><collection>Biotechnology Research Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><jtitle>Pharmaceutical research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Nielsen, Ebbe J. B</au><au>Nielsen, Jan M</au><au>Becker, Daniel</au><au>Karlas, Alexander</au><au>Prakash, Hridayesh</au><au>Glud, Sys Z</au><au>Merrison, Jonathan</au><au>Besenbacher, Flemming</au><au>Meyer, Thomas F</au><au>Kjems, Jørgen</au><au>Howard, Kenneth A</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Pulmonary Gene Silencing in Transgenic EGFP Mice Using Aerosolised Chitosan/siRNA Nanoparticles</atitle><jtitle>Pharmaceutical research</jtitle><stitle>Pharm Res</stitle><addtitle>Pharm Res</addtitle><date>2010-12-01</date><risdate>2010</risdate><volume>27</volume><issue>12</issue><spage>2520</spage><epage>2527</epage><pages>2520-2527</pages><issn>0724-8741</issn><eissn>1573-904X</eissn><abstract>Purpose This work describes the production and application of an aerosolised formulation of chitosan nanoparticles for improved pulmonary siRNA delivery and gene silencing in mice. Methods Aerosolised chitosan/siRNA nanoparticles were pneumatically formed using a nebulising catheter and sized by laser diffraction. In vitro silencing of aerosolised and non-aerosolised formulations was evaluated in an EGFP endogenous-expressing H1299 cell line by flow cytometry. Non-invasive intratracheal insertion of the catheter was used to study nanoparticle deposition by histological detection of Cy3-labeled siRNA and gene silencing in transgenic EGFP mouse lungs using a flow cytometric method. Results Flow cytometric analysis demonstrated minimal alteration in gene silencing efficiency before (68%) and after (62%) aerosolisation in EGFP-expressing H1299 cells. Intratracheal catheter administration in mice resulted in nanoparticle deposition throughout the entire lung in both alveoli and bronchiolar regions using low amounts of siRNA. Transgenic EGFP mice dosed with the aerosolised nanoparticle formulation showed significant EGFP gene silencing (68% reduction compared to mismatch group). Conclusions This work provides a technology platform for effective pulmonary delivery and gene silencing of RNAi therapeutics with potential use in preclinical studies of respiratory disease treatment.</abstract><cop>Boston</cop><pub>Boston : Springer US</pub><pmid>20824309</pmid><doi>10.1007/s11095-010-0255-y</doi><tpages>8</tpages></addata></record> |
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| subjects | aerosol Aerosols Alveoli Animals Base Sequence Biochemistry Biomedical and Life Sciences Biomedical Engineering and Bioengineering Biomedical materials Biomedicine Catheters Cell Line Chitosan Diffraction Flow Cytometry Gene Knockdown Techniques Gene Silencing Gene therapy Green Fluorescent Proteins - genetics Lasers Lung Medical Law Mice Mice, Inbred C57BL Mice, Transgenic Nanoparticles Pharmacology/Toxicology Pharmacy pulmonary gene silencing Research Paper Ribonucleic acid RNA RNA Interference RNA-mediated interference Rodents siRNA Trachea Transgenic animals Transgenic mice |
| title | Pulmonary Gene Silencing in Transgenic EGFP Mice Using Aerosolised Chitosan/siRNA Nanoparticles |
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