A novel reporter construct for screening small molecule inhibitors that specifically target self-renewing cancer cells

Shanmugam, Geetha; Mohan, Amrutha; Kumari, Khushbu; Louis, Jiss Maria; Krishnan, Usha; Balagopal, P.G.; George, Nebu Abraham; Stolzmann, Paul; Maliekal, Tessy Thomas

doi:10.1016/j.yexcr.2019.111551

Cited by 11 publications

(21 citation statements)

References 49 publications

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“…Hence, there is an increasing demand for new drugs for CSC-targeted therapy. In this context, we propose that CSCtargeting drug screening platforms can be developed using reporters of CSCs, which have been recently published (38)(39)(40)(41)(42)(43)(44)(45). These fluorescent reporters can be easily adapted for large scale screening.…”

Section: Clinical Trials For Targeting Cscsmentioning

confidence: 99%

“…Since majority of the solid tumors express ALDH1A1 in the CSC population, reporter constructs for CSCs based on ALDH1A1 promoter have been reported (40,45). A reporter construct where tdTomato is driven by ALDH1A1 promoter was used to evaluate drug sensitivity in breast cancer and colon cancer cell lines (40).…”

Section: Aldh1a1 Reportermentioning

confidence: 99%

“…Further they applied nanotechnology to this reporter system to generate a CSC model to evaluate drug efficiency to target CSCs as a co-culture system with polymer micelles loaded with chemotherapeutic drugs (40). Recently we reported a construct, ALDH1A1-DsRed2, marking CSCs in oral cancer cells to screen inhibitors for their efficacy in targeting CSCs (45). The biochemical analysis coupled with the cytotoxicity evaluation revealed the existence of a signaling cross-talk of several pathways that counteract the effect of one particular pathway inhibitor (45).…”

Section: Aldh1a1 Reportermentioning

confidence: 99%

“…Recently we reported a construct, ALDH1A1-DsRed2, marking CSCs in oral cancer cells to screen inhibitors for their efficacy in targeting CSCs (45). The biochemical analysis coupled with the cytotoxicity evaluation revealed the existence of a signaling cross-talk of several pathways that counteract the effect of one particular pathway inhibitor (45). Thus, it is evident that the outcome of pathway inhibitors predicted by RNAi and overexpression systems may not translate well in therapeutic targeting, suggesting the need to develop in vitro an in vivo drug screening platforms to target CSCs prior to clinical evaluation.…”

Section: Aldh1a1 Reportermentioning

confidence: 99%

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Reporters of Cancer Stem Cells as a Tool for Drug Discovery

Mohan

et al. 2021

Front. Oncol.

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In view of the importance of cancer stem cells (CSCs) in chemoresistance, metastasis and recurrence, the biology of CSCs were explored in detail. Based on that, several modalities were proposed to target them. In spite of the several clinical trials, a successful CSC-targeting drug is yet to be identified. The number of molecules screened and entered for clinical trial for CSC-targeting is comparatively low, compared to other drugs. The bottle neck is the lack of a high-throughput adaptable screening strategy for CSCs. This review is aimed to identify suitable reporters for CSCs that can be used to identify the heterogeneous CSC populations, including quiescent CSCs, proliferative CSCs, drug resistant CSCs and metastatic CSCs. Analysis of the tumor microenvironment regulating CSCs revealed that the factors in CSC-niche activates effector molecules that function as CSC markers, including pluripotency markers, CD133, ABCG2 and ALDH1A1. Among these factors OCT4, SOX2, NANOG, ABCG2 and ALDH1A1 are ideal for making reporters for CSCs. The pluripotency molecules, like OCT4, SOX2 and NANOG, regulate self-renewal, chemoresistance and metastasis. ABCG2 is a known regulator of drug resistance while ALDH1A1 modulates self-renewal, chemoresistance and metastasis. Considering the heterogeneity of CSCs, including a quiescent population and a proliferative population with metastatic ability, we propose the use of a combination of reporters. A dual reporter consisting of a pluripotency marker and a marker like ALDH1A1 will be useful in screening drugs that target CSCs.

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Section: Clinical Trials For Targeting Cscsmentioning

confidence: 99%

Section: Aldh1a1 Reportermentioning

confidence: 99%

Section: Aldh1a1 Reportermentioning

confidence: 99%

Section: Aldh1a1 Reportermentioning

confidence: 99%

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Reporters of Cancer Stem Cells as a Tool for Drug Discovery

Mohan

et al. 2021

Front. Oncol.

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“…Thus even in a single tumor, there could be heterogeneity in the CSCs, depending on the niche they reside ( Visvader and Lindeman, 2012 ). The existence of a dormant CSC population as well as proliferative CSCs is known in many cancers, and they show different levels of differentiation ( Bliss et al, 2016 ; Shanmugam et al, 2019 ). Although CSCs might have properties like tumor initiation capacity, drug resistance, and/or metastatic ability, a single CSC at a given time point may not show all the three properties.…”

Section: Introductionmentioning

confidence: 99%

Markers and Reporters to Reveal the Hierarchy in Heterogeneous Cancer Stem Cells

Mohan

Rajan

Mohan

et al. 2021

Front. Cell Dev. Biol.

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A subpopulation within cancer, known as cancer stem cells (CSCs), regulates tumor initiation, chemoresistance, and metastasis. At a closer look, CSCs show functional heterogeneity and hierarchical organization. The present review is an attempt to assign marker profiles to define the functional heterogeneity and hierarchical organization of CSCs, based on a series of single-cell analyses. The evidences show that analogous to stem cell hierarchy, self-renewing Quiescent CSCs give rise to the Progenitor CSCs with limited proliferative capacity, and later to a Progenitor-like CSCs, which differentiates to Proliferating non-CSCs. Functionally, the CSCs can be tumor-initiating cells (TICs), drug-resistant CSCs, or metastasis initiating cells (MICs). Although there are certain marker profiles used to identify CSCs of different cancers, molecules like CD44, CD133, ALDH1A1, ABCG2, and pluripotency markers [Octamer binding transcriptional factor 4 (OCT4), SOX2, and NANOG] are used to mark CSCs of a wide range of cancers, ranging from hematological malignancies to solid tumors. Our analysis of the recent reports showed that a combination of these markers can demarcate the heterogeneous CSCs in solid tumors. Reporter constructs are widely used for easy identification and quantification of marker molecules. In this review, we discuss the suitability of reporters for the widely used CSC markers that can define the heterogeneous CSCs. Since the CSC-specific functions of CD44 and CD133 are regulated at the post-translational level, we do not recommend the reporters for these molecules for the detection of CSCs. A promoter-based reporter for ABCG2 may also be not relevant in CSCs, as the expression of the molecule in cancer is mainly regulated by promoter demethylation. In this context, a dual reporter consisting of one of the pluripotency markers and ALDH1A1 will be useful in marking the heterogeneous CSCs. This system can be easily adapted to high-throughput platforms to screen drugs for eliminating CSCs.

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