In-vivo Single Cell Transcript Analyses for Systems Modelling
Conference Recording Jun 12, 2014
About the Speaker
Philip Day is Reader in Quantitative Analytical Genomics, Manchester University, where he develops innovative tools for accurate measurement of transcripts in heterogeneous tissues for application in cancer studies. Philip heads a group investigating the systems pathology of neuroblastoma and leukaemia, and molecular biomarkers relating to prognostics and function. Current studies focus on assay miniaturization for single cell analyses, enabling drug response measurement across cell populations and cell-cell interactions. He is engaging biomarker integration and modelling for enabling personalised healthcare and is developing gene synthesis platforms for discovery in synthetic biology. He has over 100 peer reviewed papers, is an examiner for various research councils and journals, and is an RSC Editorial Board member. Abstract
To investigate the importance of minimal residual disease of BCR-ABL expressing cells chronic myelogenous leukemia (CML) was assessed using single cell molecular analytical approaches. CML is believed to occur as a consequence of the clonal expansion of leukemic stem cells and to be maintained by an expanding population of hematopoietic stem cells that have acquired a BCR-ABL fusion gene. Recent studies indicate that primitive CML cells are less responsive to tyrosine kinase inhibitors and are a reservoir for the emergence of tyrosine kinase resistant subclones. It also has been reported that BCR-ABL mediated cell adhesion may be involved in post-therapy residual disease of CML. The natural heterogeneity seen across both attached and unattached BCR-ABL expressing populations has been investigated. The study developed a means to utilize flow-assisted cell sorting of cell lines expressing BCR-ABL to derive individual attached and unattached cell sub-populations. Using a homogenous extraction procedure, cells individually flow sorted into microtitre plates were subjected to combinations of ABL and BCR-ABL RT qPCR, and also phosphorylated BCR-ABL protein was assessed using the proximity ligation assay and revealed the existence of lowly and highly BCR-ABL expressing cell line populations. The implications of this study will be discussed along with newly derived systems models and in-vivo sample procedures employing AFM.