High Throughput Single-Cell Phenotype Isolation by Protrusion Analysis Chip (PAC) There is increasing evidence that solid tumors are likely comprised of many subpopulations of cells with distinct genotypes and phenotypes, which is a phenomenon termed intratumor heterogeneity. Such heterogeneity becomes a major obstacle to effective cancer treatment and personalized medicine. Because of this inherent heterogeneity, data collected from cancer cell population-averaged assays likely hides valuable but rare events such as dramatic variations in gene expression at the single cell level. Therefore, understanding cellular heterogeneity from cancer biospecimens, especially in the study of phenotype-genotype correlation, will facilitate identification of new cell subsets, and assist in cancer prevention, diagnosis, and therapy. In this proposal, we focus on developing a high throughput approach for single-cell isolation based on cells' capability in generating protrusions. To be able to retrieve the desired single adherent cell, a Protrusion Analysis Chip (PAC) is proposed, in which the single cell is captured by a single hook and physically isolated by a barrier. The PAC is rapid, operationally simple, highly efficient, and requires low-volume sample introduction. After adhesion and spreading, cell phenotype is identified microscopically and then the desired cell is retrieved for genotype analysis. The proposed technology developments and study plans may potentially strengthen the understanding of the relationship between phenotype and genotype at the single cell level.

Public Health Relevance

High Throughput Single-Cell Phenotype Isolation by Protrusion Analysis Chip (PAC) In this proposal, we plan to develop a new method to selectively isolate a single adherent cell with the desired phenotype for cancer heterogeneity research. The isolated cancer cells can be used for single cell analysis and biofunction-genotype correlation. Our approach may provide an efficient cell assay tool for cancer bio-specimen study.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21CA191179-03
Application #
9316554
Study Section
Special Emphasis Panel (ZCA1)
Program Officer
Knowlton, John R
Project Start
2015-08-15
Project End
2018-07-31
Budget Start
2017-08-01
Budget End
2018-07-31
Support Year
3
Fiscal Year
2017
Total Cost
Indirect Cost
Name
Methodist Hospital Research Institute
Department
Type
DUNS #
185641052
City
Houston
State
TX
Country
United States
Zip Code
77030
Vu, Timothy Quang; de Castro, Ricardo Miguel Bessa; Qin, Lidong (2017) Bridging the gap: microfluidic devices for short and long distance cell-cell communication. Lab Chip 17:1009-1023
Liu, Zongbin; Han, Xin; Zhou, Qing et al. (2017) Integrated Microfluidic System for Gene Silencing and Cell Migration. Adv Biosyst 1:
Li, Ying; Uddayasankar, Uvaraj; He, Bangshun et al. (2017) Fast, Sensitive, and Quantitative Point-of-Care Platform for the Assessment of Drugs of Abuse in Urine, Serum, and Whole Blood. Anal Chem 89:8273-8281
Ma, Yuan; Han, Xin; Quintana Bustamante, Oscar et al. (2017) Highly efficient genome editing of human hematopoietic stem cells via a nano-silicon-blade delivery approach. Integr Biol (Camb) 9:548-554
Jo, Myeong Chan; Qin, Lidong (2016) Microfluidic Platforms for Yeast-Based Aging Studies. Small 12:5787-5801
Zhang, Kai; Gao, Min; Chong, Zechen et al. (2016) Single-cell isolation by a modular single-cell pipette for RNA-sequencing. Lab Chip 16:4742-4748
Han, Xin; Liu, Zongbin; Zhao, Li et al. (2016) Microfluidic Cell Deformability Assay for Rapid and Efficient Kinase Screening with the CRISPR-Cas9 System. Angew Chem Int Ed Engl 55:8561-5
Liu, Zongbin; Han, Xin; Qin, Lidong (2016) Recent Progress of Microfluidics in Translational Applications. Adv Healthc Mater 5:871-88