Conditional Reprogramming for Patient-Derived Cancer Models and Next-Generation Living Biobanks

Abstract

Traditional cancer models including cell lines and animal models have limited applications in both basic and clinical cancer research. Genomics-based precision oncology only help 2–20% patients with solid cancer. Functional diagnostics and patient-derived cancer models are needed for precision cancer biology. In this review, we will summarize applications of conditional cell reprogramming (CR) in cancer research and next generation living biobanks (NGLB). Together with organoids, CR has been cited in two NCI (National Cancer Institute, USA) programs (PDMR: patient-derived cancer model repository; HCMI: human cancer model initiatives. HCMI will be distributed through ATCC). Briefly, the CR method is a simple co-culture technology with a Rho kinase inhibitor, Y-27632, in combination with fibroblast feeder cells, which allows us to rapidly expand both normal and malignant epithelial cells from diverse anatomic sites and mammalian species and does not require transfection with exogenous viral or cellular genes. Establishment of CR cells from both normal and tumor tissue is highly efficient. The robust nature of the technique is exemplified by the ability to produce 2 × 106 cells in five days from a core biopsy of tumor tissue. Normal CR cell cultures retain a normal karyotype and differentiation potential and CR cells derived from tumors retain their tumorigenic phenotype. CR also allows us to enrich cancer cells from urine (for bladder cancer), blood (for prostate cancer), and pleural effusion (for non-small cell lung carcinoma). The ability to produce inexhaustible cell populations using CR technology from small biopsies and cryopreserved specimens has the potential to transform biobanking repositories (NGLB: next-generation living biobank) and current pathology practice by enabling genetic, biochemical, metabolomic, proteomic, and biological assays, including chemosensitivity testing as a functional diagnostics tool for precision cancer medicine. We discussed analyses of patient-derived matched normal and tumor models using a case with tongue squamous cell carcinoma as an example. Last, we summarized applications in cancer research, disease modeling, drug discovery, and regenerative medicine of CR-based NGLB.

Patient-Derived In Vitro Models for Drug Discovery in Colorectal Carcinoma

“…Y27632 has been shown to induce the proliferative capacity of primary tumor cells resulting in efficient immortalization of cells into stratified epithelium without any DNA damage [37,38]. Several groups have elaborated specific protocols for cell isolation from various tumor types, including hepatocellular carcinoma [39], prostate cancer [40], tongue squamous cell carcinoma [41] and non-small lung cancers [42]. Liu et al described a detailed protocol of CRC patient-derived cell cultures establishment from both cancerous and non-cancerous tissue biopsies that had the capacity to grow indefinitely in vitro, while maintaining phenotypic and genotypic features of the original tissue [36].…”

Personalized drug testing in a patient with non-small-cell lung cancer using cultured cancer cells from pleural effusion

“…[5][6][7] Conditional reprogramming (CR) is a new technique for rapidly expanding normal and tumor cells from surgical specimens or needle biopsies. 8,9 CR technology has displayed promise in basic and clinical cancer research, 10 especially for the rapid screening of candidate drugs and personalized medicine. [11][12][13] The genomic and histological features of the original tumor are maintained in cultured CR cells.…”

Conditionally Reprogrammed Human Normal Airway Epithelial Cells at ALI: A Physiological Model for Emerging Viruses

“…The viral tropism correlates to expression of viral receptor(s) on the surface of host cells and intracellular restriction as well. Cancer cell lines have been used widely for biological or functional cell systems in many other fields (Agarwal and Rimm 2012;Barretina et al 2012;Palechor-Ceron et al 2019). These cells are defective for many functions of normal human cells because of significant genetic and epigenetic changes compared to the normal tissues.…”

“…These cell cultures stop proliferating if one of the conditions (Y-27632 or feeder layer) is not met. We termed this culture method and resulting cells as ''conditional reprogramming (CR)'' and ''conditionally reprogrammed cells (CRC)'', respectively (Liu et al 2012;Suprynowicz et al 2012;Palechor-Ceron et al 2013;Liu et al 2017;Palechor-Ceron et al 2019). Indeed, organoids (Weeber et al 2017;Puca et al 2018;Sachs et al 2018;Mullenders et al 2019;Xinaris 2019) and CR technologies have been widely used in cancer biology and regeneration fields.…”

“…Comparison of ex vivo cell models.CRCs/ALI: An Ex Vivo Models for Virus InfectionsTo date, CR by itself or in combination with ALI cultures have been used in studies of host defense and viral infections, drug screening and toxicity testing, wound healing/repair, gene therapies as ex vivo preclinical models for lung cancer, chronic obstructive pulmonary disease, cystic fibrosis, and asthma(Yuan et al 2012;Bove et al 2014;Crystal et al 2014;Saenz et al 2014; Chu et al 2015b;Feng et al 2015;Kotha et al 2015;Walters et al 2015;Alamri et al 2016;Beglyarova et al 2016;Butler et al 2016;Ellis et al 2016;Papapetrou 2016;Reynolds et al 2016; Borodovsky et al Brewington et al 2018;Correa et al 2018;Jin et al 2018;LaRanger et al 2018;Mimoto et al 2018;Moorefield et al 2018;Peters-Hall et al 2018;Shay et al 2018;Wang et al 2018; Zhang Z et al 2018;Alkhilaiwi et al 2019;Jiang et al 2019;Krawczyk et al 2020;Martini et al 2019;Mondal et al 2019;Nicolas et al 2019;Palechor-Ceron et al 2019;Su et al 2019;Chai et al 2020). Compared to organoids (close 3D cultures) shown in…”