dc.contributor.author | Hayman, Melissa Anne | |
dc.date.accessioned | 2018-04-04T15:45:59Z | |
dc.date.available | 2018-04-04T15:45:59Z | |
dc.date.issued | 2018-03-19 | |
dc.date.submitted | 2018-04-04T13:47:28.309Z | |
dc.identifier.citation | Hayman, M.A. 2018. Genomic influences on platelet function. Genomic influences on platelet function | en_US |
dc.identifier.uri | http://qmro.qmul.ac.uk/xmlui/handle/123456789/36221 | |
dc.description | PhD | en_US |
dc.description.abstract | The study of platelet messenger and micro-RNAs is of increasing interest
owing to the fact that platelets contain the machinery to splice and
translate mRNA into proteins in response to inhibitory or activating
signals. However, the relatively small size (roughly 4000-5000 transcripts)
and short half-life of the platelet transcriptome makes this a technically
challenging aspect of platelet biology to investigate.
The aims of these thesis investigations were therefore to optimise
protocols for the isolation of platelets for downstream RNA analyses and
function testing, to investigate the functional capabilities of platelet
subpopulations rich in RNA, and to understand the functional and
transcriptomic impact of gene mutations predicted to influence platelet
function.
I found that the optimal method for isolating platelets from whole blood is
to use simple single step centrifugation to obtain platelet rich plasma.
This method is as effective as more involved methods at reducing white
blood cell contamination whilst causing minimal platelet activation. Using
this method in combination with flow cytometric cell sorting techniques I
was able to isolate the newly formed reticulated platelet sub-population
and to confirm the link between reticulation status and increased RNA
content. Furthermore, using a range of platelet function assays I
demonstrated that reticulated platelets are more reactive than
non-reticulated platelets. By obtaining blood samples from a patient with
a PLA2G4A mutation I was able to show that loss of cPLA2α enzymatic
activity alters both platelet function and the expression of certain mRNA
transcripts. My investigations using samples from a range of patients with
bleeding tendencies show the benefit of combining deep platelet
phenotyping with next generation sequencing to understand the
causation of bleeding disorders.
Together these investigations highlight the utility of genomic DNA and
platelet specific mRNA studies in providing novel insights in to pathways
regulating platelet reactivity. | en_US |
dc.language.iso | en | en_US |
dc.publisher | Genomic influences on platelet function | en_US |
dc.rights | The copyright of this thesis rests with the author and no quotation from it or information derived from it may be published without the prior written consent of the author | |
dc.subject | micro-RNAs | en_US |
dc.subject | platelets | en_US |
dc.subject | platelet biology | en_US |
dc.title | Genomic influences on platelet function | en_US |
dc.type | Thesis | en_US |