It is clear from numerous reports to date that the involvement of miRNA in the pathology of CF plays a major role. The specific research question here however is a little unclear as the proposal appears to ask two different research questions, firstly, (Tasks 1-4) the aim is to examine pro inflammatory miRNA and their targets with a view to managing inflammatory associated tissue destruction in CF and secondly (Tasks 5-8) is to explore miRNA and CFTR function. The investigators try to link these two aims however the project is very ambitious for a two year period. It could easily be separated into two different proposals. If delivered upon though it will greatly extend the current body of work in this area providing new leads for the management of IL-8 induced inflammation in CF known to potentiate tissue destruction in this condition. The use of PNA’s is a novel approach and may indeed uncover potential therapeutic targets. The objectives of this proposal MICRORNA-CF are listed as: (a) identification of microRNAs which are either down- or up-regulated in association with P.aeruginosa infection of CF bronchial epithelial cells; (b) identification and validation of the most relevant mRNA targets of the modulated microRNAs; (c) study of changes of the gene expression and secretome profile of CF using PNAs targeting microRNAs; (d) development and biological validation of novel PNA-based molecules interfering with microRNAs involved in inflammation; (e) development of PNA-based antagomiRNAs for CFTR stabilization. Overall it is a really nice project and worthwhile to add to what is currently known about miRNA in the progression of CF and the battle to find new ways to treat this condition but I’m cautious as to the amount of work to be performed in a two year period. Nonetheless the investigators have a proven track record and with their team of collaborators with the experience detailed this may be possible. The use of PNA’s and tasks 5-7 in the experimental plan is very interesting especially the planned testing with CFTR correctors. Contingency plans should be in place for the possibility that miRNA and their targets identified in immortilised cell lines may not be the same as those observed in primary cells. Transfection of polarised cells can be a difficult process as the cell density required to maintain TEER are very unfavourable for transfection of pre and antimiR’s, this may not be the case for PNA’s however. Again results on polarised cells may not exactly reflect those observed in monolayers yet this is an important part of the experimental plan. If the investigators could gain access to clinical samples it would greatly strengthen their work. I’m not sure about the primary cell model – is this a mix of normal and CF bronchial epithelial cells? Task 3 (experimental plan) – consideration should also be given to those miRs decreased by PA01 treatment as subsequent increased expression of their targets may be pro-inflammatory.
MicroRNA Therapeutics in CF: Targeting CFTR and inflammation networks (MICRORNA-CF)”
Gambari R.
2016
Abstract
It is clear from numerous reports to date that the involvement of miRNA in the pathology of CF plays a major role. The specific research question here however is a little unclear as the proposal appears to ask two different research questions, firstly, (Tasks 1-4) the aim is to examine pro inflammatory miRNA and their targets with a view to managing inflammatory associated tissue destruction in CF and secondly (Tasks 5-8) is to explore miRNA and CFTR function. The investigators try to link these two aims however the project is very ambitious for a two year period. It could easily be separated into two different proposals. If delivered upon though it will greatly extend the current body of work in this area providing new leads for the management of IL-8 induced inflammation in CF known to potentiate tissue destruction in this condition. The use of PNA’s is a novel approach and may indeed uncover potential therapeutic targets. The objectives of this proposal MICRORNA-CF are listed as: (a) identification of microRNAs which are either down- or up-regulated in association with P.aeruginosa infection of CF bronchial epithelial cells; (b) identification and validation of the most relevant mRNA targets of the modulated microRNAs; (c) study of changes of the gene expression and secretome profile of CF using PNAs targeting microRNAs; (d) development and biological validation of novel PNA-based molecules interfering with microRNAs involved in inflammation; (e) development of PNA-based antagomiRNAs for CFTR stabilization. Overall it is a really nice project and worthwhile to add to what is currently known about miRNA in the progression of CF and the battle to find new ways to treat this condition but I’m cautious as to the amount of work to be performed in a two year period. Nonetheless the investigators have a proven track record and with their team of collaborators with the experience detailed this may be possible. The use of PNA’s and tasks 5-7 in the experimental plan is very interesting especially the planned testing with CFTR correctors. Contingency plans should be in place for the possibility that miRNA and their targets identified in immortilised cell lines may not be the same as those observed in primary cells. Transfection of polarised cells can be a difficult process as the cell density required to maintain TEER are very unfavourable for transfection of pre and antimiR’s, this may not be the case for PNA’s however. Again results on polarised cells may not exactly reflect those observed in monolayers yet this is an important part of the experimental plan. If the investigators could gain access to clinical samples it would greatly strengthen their work. I’m not sure about the primary cell model – is this a mix of normal and CF bronchial epithelial cells? Task 3 (experimental plan) – consideration should also be given to those miRs decreased by PA01 treatment as subsequent increased expression of their targets may be pro-inflammatory.I documenti in SFERA sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
