TOPIC 382 - INTEGRATED IMAGING AND MOLECULAR ANALYSIS OF GLIOBLASTOMA USING SCOPE SEQ ON THE AIR FLOW

Clinical research is incorporating single cell analysis methods into various evaluations.

During 75N91019C00029 (Phase I) of this proposed program, we intended to validate the performance of a novel single cell transcriptomic (i.e. sc-RNA-Seq) sample preparation platform which integrates imaging into the overall workflow.

SCOPE-Seq on the AIR FLOW System will link imaging data to sc-RNA-Seq data on a cell by-cell basis allowing clinical outcomes such as resection margins and therapeutic success to be linked to a cell?s transcriptomic profile.

Here, we will acquire primary biopsy samples from glioblastoma patients undergoing surgical resection during a clinical evaluation of dyes used in fluorescent-guided surgery. One of the dyes, 5-ALA, has been reported to preferentially label tumor cells over non-disease subpopulations.

Patient-derived cells will be processed on Cell Microsystems' AIR FLOW System using the novel Single Cell Optical Phenotyping and Expression Sequencing (SCOPESeq) method.

Using SCOPE-Seq?s optical barcoding methods on the automated AIR FLOW System, we will link the imaging data (e.g. 5-ALA positive vs. negative) with sc-RNA-Seq data on a cell-by-cell basis.

While not part of the research endpoints, the resulting data set is intended to determine tumor-cell staining specificity of dyes while using sc-RNA-Seq analysis to reveal cellular subpopulations with high resolution.

The goal of the program is to examine clinically derived glioma cells for presence of the fluorescent dye 5-ALA which is used as a visual guide during tumor resection.

The AIR FLOW System will be used to determine which cells are 5-ALA positive and which exhibit transcriptomic properties of glioma cells as a means of testing the sensitivity and specificity of 5-ALA as a visual guide during surgery.

The proposal also seeks to validate the use of the CITE-Seq/TotalSeq workflow, a means of using oligo-tagged antibodies for protein quantification using next generation sequencing, in the overall glioma phenotyping assay.