Luminescent phospholipid analogs for the identification and isolation of circulating tumor cells
Inventors
Weichert, Jamey P. • Pak, Chorom • PINCHUK, Anatoly • Kozak, Kevin • LONGINO, Marc
Assignees
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Publication Number
US-11467159-B2
Publication Date
2022-10-11
Expiration Date
Abstract
The present invention is directed to a method of identifying, isolating, and enabling downstream analysis of circulating tumor cells comprising contacting a blood or blood serum sample of a subject with a composition comprising a phospholipid ether analog bound to a luminescent molecule or a magnetic bead and subjecting the blood or blood serum sample of the subject to fluorescent microscopy, flow cytometry or magnetic isolation.
Core Innovation
The invention relates to identifying and isolating circulating tumor cells (CTCs) by contacting a blood or blood serum sample from a subject having one or more circulating tumor cells with a compound of formula (I). The compound of formula (I) includes an integer n from 16 to 30, Y selected from —H, —OH, —OR, —C(O)OH, and —OC(O)R, and X as a luminescent molecule. After contacting, the blood or blood serum sample is subjected to flow cytometry.
After flow cytometry, the one or more CTCs are isolated based on the luminescent compound of formula (I). The isolated CTCs are selected from a group that includes breast cancer cells, lung cancer cells, thyroid cancer cells, cervical cancer cells, squamous cell carcinoma cells, carcinoma cells, prostate cancer cells, pancreas cancer cells, colorectal cancer cells, and cancer stem cells.
The described approach is supported by a mechanistic rationale connected to lipid rafts and cholesterol affinity, using phospholipid ether (PLE) analogs. In representative results described in the document, a fluorescent PLE analog (CLR1501) is used for flow-cytometry marker uptake and quantitative enumeration across multiple cancer types, including comparisons against conventional CTC definitions based on EpCAM/CK.
Claims Coverage
Independent claim coverage centers on three inventive features: contacting a blood or blood serum sample with a specified luminescent PLE-analog compound of formula (I), performing flow cytometry, and isolating CTCs selected from multiple cancer cell types including cancer stem cells.
Contacting blood or blood serum with a luminescent compound of formula (I)
Contacting a blood or blood serum sample from a subject having one or more circulating tumor cells with a compound of formula (I) where n is an integer from 16 to 30, Y is selected from —H, —OH, —OR, —C(O)OH, and —OC(O)R, and X is a luminescent molecule.
Flow cytometry of the contacted blood or serum sample
Subjecting the blood or blood serum sample to flow cytometry after contacting with the compound of formula (I).
Isolating CTCs selected from multiple cancer cell types
Isolating the one or more CTCs following flow cytometry, where the one or more CTCs are selected from the group consisting of a breast cancer cell, a lung cancer cell, a thyroid cancer cell, a cervical cancer cell, a squamous cell, a carcinoma cell, a prostate cancer cell, a pancreas cancer, a colorectal cancer cell, and a cancer stem cell.
Using a fluorophore as the luminescent molecule
The luminescent molecule X is a fluorophore.
Constrained fluorophore substituent R groups
Selecting the fluorophore such that each R substituent on the fluorophore is independently H, CH3, C2H5, or C3H7.
Flow cytometry is fluorescence-activated cell sorting
The flow cytometry step is fluorescence-activated cell sorting.
Using isolated CTCs for specified molecular and cellular analyses
Using one or more isolated CTCs in a selected technology chosen from protein isolation, RNA isolation, DNA isolation, gene translocation analysis, gene amplification analysis, and fluorescent in-situ hybridization.
Prostate cancer cell specificity
The circulating tumor cells are prostate cancer cells.
The claim coverage centers on isolating CTCs using a specified phospholipid ether (PLE) analog of formula (I) with a luminescent molecule X, followed by flow cytometry and CTC isolation. Dependent claims narrow the luminescent molecule to specific fluorophores and constraining substituents, specify fluorescence-activated cell sorting, identify downstream analyses, and further narrow the CTCs to prostate cancer cells.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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