Patent ID: 9220793
Filing Date: 2015-12-29
Classification: A61K,C07H,C12Q

Abstract:
1. A method for in vivo identification of remnant living cells in late-stage tumors as cancer initiating cells (CICs) in a living rodent animal, comprising steps of: (a) providing a multimodality reporter gene imaging system for co-expressing multiple reporter genes, wherein the multimodality reporter gene imaging system is delivered by a piggyBac transposon system comprising a helper plasmid that encodes the piggyBac transposase gene, and a donor plasmid comprising a constitutive gene promoter and two reporter genes for in vivo imaging the transcription and translation activity of genes cloned at the downstream of the constitutive gene promoter and an antibiotic-resistant cassette for transposition assay and stable clone selection; (b) co-transfecting the donor plasmid comprising the two reporter genes and the helper plasmid into a cancer cell line; (c) implanting the transfected cancer cells into a living rodent animal by subcutaneous (SC) inoculation; (d) tracking the tumor progression and location of remnant living cancer cells that escape loss or death in a late-stage tumor bearing in the rodent animal by the multimodality reporter gene imaging; (e) isolating the remnant living cells that escape cell loss or death in a late-stage tumor for an assay of sphere formation or cancer initiating cell biomarker genes; and (f) identifying the remnant living cells that escape cell loss or death in a late-stage tumor as candidate cancer initiating cells —when the amount of sphere formation and cancer initiating cell biomarker genes increase.