Sparks et al, Heterogeneity in tumor chromatin-doxorubicin binding revealed by in vivo fluorescence lifetime imaging confocal endomicroscopy: MATLAB code

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Title: Sparks et al, Heterogeneity in tumor chromatin-doxorubicin binding revealed by in vivo fluorescence lifetime imaging confocal endomicroscopy: MATLAB code
Authors: Sparks, H
Munro, I
Warren, S
Item Type: Software / Code
Abstract: Abstract We present an approach to quantify drug-target engagement using in vivo fluorescence endomicroscopy, validated with in vitro measurements. Specifically, we demonstrate that doxorubicin binding to chromatin causes a change in the fluorescence lifetime of histone-GFP fusions that can be measured in vivo at single cell resolution using a confocal laparo/endomicroscope. We measure both intra- and inter-tumor heterogeneity in doxorubicin chromatin engagement in a model of peritoneal metastasis of ovarian cancer, revealing striking variation in the efficacy of doxorubicin-chromatin binding depending on intra-peritoneal or intra-venous delivery. In addition, we observe significant variations in doxorubicin-chromatin binding between different metastases in the same mouse and between different regions of the same metastasis. The quantitative nature of fluorescence lifetime imaging enables direct comparison of drug target engagement for different drug delivery modalities and between in vitro and in vivo experiments. This uncovers dramatically different rates of cell killing for the same level of doxorubicin binding in vitro and in vivo.
Abstract We present an approach to quantify drug-target engagement using in vivo fluorescence endomicroscopy, validated with in vitro measurements. Specifically, we demonstrate that doxorubicin binding to chromatin causes a change in the fluorescence lifetime of histone-GFP fusions that can be measured in vivo at single cell resolution using a confocal laparo/endomicroscope. We measure both intra- and inter-tumor heterogeneity in doxorubicin chromatin engagement in a model of peritoneal metastasis of ovarian cancer, revealing striking variation in the efficacy of doxorubicin-chromatin binding depending on intra-peritoneal or intra-venous delivery. In addition, we observe significant variations in doxorubicin-chromatin binding between different metastases in the same mouse and between different regions of the same metastasis. The quantitative nature of fluorescence lifetime imaging enables direct comparison of drug target engagement for different drug delivery modalities and between in vitro and in vivo experiments. This uncovers dramatically different rates of cell killing for the same level of doxorubicin binding in vitro and in vivo.
Issue Date: 17-May-2018
URI: http://hdl.handle.net/10044/1/64505
DOI: https://doi.org/10.5281/zenodo.1249004
Copyright Statement: https://creativecommons.org/licenses/by-sa/4.0/
Keywords: Sparks et al, Heterogeneity in tumor chromatin-doxorubicin binding revealed by in vivo fluorescence lifetime imaging confocal endomicroscopy
Notes: MATLAB code: takes as input FIFO TCSPC frames in .pff file format generated by the FLIM confocal endomicroscope (CEM) converts from .pff format to .raw format for manipulation of TCSPC data as a matrix in MATLAB uses a template matching algorithm to align FIFO TCSPC framee and combines aligned frames into a single integrated FLIM image stored as a series of .Tif image generates cell nuclei binary segmentation masks for FLIMfit converts background and reference fluorophore data in form of .pff into FLIM image stored as a series of .Tif image
Appears in Collections:Faculty of Natural Sciences - Research Data



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