Type

Journal Article

Authors

Sally-Ann Cryan
Joanne M Ramsey
Neeraj Sivadas
Noel G McElvaney
Catherine M Greene
Joanne O'Dwyer
Katie Nolan
Ciaran Lawlor
Awadh B Yadav
Ciara Kelly

Subjects

Biochemistry

Topics
cell line poly lactic acid laser scanning lung disease gene expression flow cytometry high content analysis gene silencing

Therapeutic aerosol bioengineering of siRNA for the treatment of inflammatory lung disease by TNFα gene silencing in macrophages. (2014)

Abstract The development of small interfering RNA (siRNA) to silence specific genes offers a new means of understanding and treating a range of respiratory diseases, including inflammatory lung disease. The alveolar macrophage (AM) is a key component of the inflammatory process in the lungs, associated with high levels of gene expression in inflammatory lung disease and therefore an attractive target for therapeutic siRNA. Delivery of siRNA to macrophages presents a significant delivery challenge, as fully differentiated alveolar macrophages are difficult to access and transfect. In this study we engineered particles suitable for inhalation that would efficiently transfect macrophages postinhalation. The process for encapsulation of siRNA in poly(lactic-co-glycolic acid) microparticles (MPs) was optimized using a double emulsion technique, and the resulting particles were characterized for size, shape, aerosol characteristics, encapsulation efficiency, and integrity of encapsulated siRNA. The cell uptake of the siRNA-loaded microparticles was determined by flow cytometry, confocal laser scanning microscopy (CLSM), and high-content analysis (HCA) with MPs capable of transfecting up to 55% of cells. Anti-TNFα siRNA-MPs were then prepared to study the functional activity of encapsulated siRNA in LPS-stimulated macrophages as a model of inflammation. The anti-TNFα siRNA-MPs were able to decrease TNFα expression by 45% over 48 h in the differentiated human monocytic cell line THP-1 compared to negligible knockdown using commercial transfection reagents and offered significant, sustained siRNA knockdown of TNFα in primary monocytes for up to 72 h.
Collections Ireland -> Royal College of Surgeons in Ireland -> PubMed

Full list of authors on original publication

Sally-Ann Cryan, Joanne M Ramsey, Neeraj Sivadas, Noel G McElvaney, Catherine M Greene, Joanne O'Dwyer, Katie Nolan, Ciaran Lawlor, Awadh B Yadav, Ciara Kelly

Experts in our system

1
Sally-Ann Cryan
Royal College of Surgeons in Ireland
Total Publications: 70
 
2
Joanne M Ramsey
Royal College of Surgeons in Ireland
Total Publications: 5
 
3
Noel G McElvaney
Royal College of Surgeons in Ireland
Total Publications: 194
 
4
Catherine M Greene
Royal College of Surgeons in Ireland
Total Publications: 150
 
5
Katie Nolan
Royal College of Surgeons in Ireland
Total Publications: 5
 
6
Ciaran Lawlor
Royal College of Surgeons in Ireland
Total Publications: 8