Intracellular protein trafficking kinetics in chronic myeloid leukemia stem cells using a microfluidic platform.

Integrative Biology : Quantitative Biosciences From Nano to Macro
Shannon L FaleyJonathan M Cooper

Abstract

The oncogenic fusion protein BCR-ABL is produced by chronic myeloid leukemia (CML) cells and functions as an abnormal, constitutively active tyrosine kinase that interferes with normal migratory and apoptotic behaviour of cells. Small molecule tyrosine kinase inhibitors (TKIs), such as dasatinib, eliminate CML progenitor cells, but fail to target the stem cell fraction resulting in persistent disease. In order to achieve a cure for CML in the majority of patients, we need an improved understanding of intracellular signalling dynamics, including the shuttling of BCR-ABL between cytosolic and nuclear compartments. In the past, the instability of BCR-ABL in assays using conventional immunohistochemical techniques has made this difficult and has not allowed for reliable analysis at the single cell level. Here we show how the utilization of rapid on-chip cell fixation within a microfluidic platform provides a means to immunofluorescently analyze the spatiotemporal localization of both BCR-ABL and c-ABL, as well as the linked apoptosis mediator, BCL-XL, in arrays of single CD34+ CML stem/progenitor cells, without cell loss. We demonstrate this proceeds up to 4 times faster than benchtop methods. Our results indicate that whilst both ...Continue Reading

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Citations

Feb 23, 2013·Advanced Drug Delivery Reviews·Xin Ting ZhengChang Ming Li
Oct 24, 2014·Lab on a Chip·Mayuree ChanasakulniyomJonathan M Cooper
Jan 1, 2018·Journal of Molecular Biology·Jérôme CharmetTuomas P J Knowles

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