A genome‐wide CRISPR activation screen reveals Hexokinase 1 as a critical factor in promoting resistance to multi‐kinase inhibitors in hepatocellular carcinoma cells

Sofer, Summer; Lamkiewicz, Kevin; Eilat, Shir Armoza; Partouche, Shirly; Marz, Manja; Moskovits, Neta; Stemmer, Salomon M.; Shlomai, Amir; Sklan, Ella H.

doi:10.1096/fj.202101507rr

Cited by 18 publications

(11 citation statements)

References 41 publications

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“…Tyrosine kinase inhibitors are considered to be the first-line therapy for patients with advanced or metastatic HCC. Of note, transcriptome analysis [ 14 ] demonstrated that TRIM15 was abnormally upregulated in regorafenib-resistant HCC cell lines (Fig. 1a ).…”

Section: Resultsmentioning

confidence: 99%

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TRIM15 forms a regulatory loop with the AKT/FOXO1 axis and LASP1 to modulate the sensitivity of HCC cells to TKIs

et al. 2023

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For patients with advanced or metastatic Hepatocellular carcinoma (HCC) who are not suitable for surgical resection, systemic therapy has been considered to be the standard treatment. In recent years, a small subset of patients with unresectable HCC have been benefit from tyrosine kinase inhibitors (TKIs), and the overall survival time of these patients is significantly increased. However, all responders ultimately develop resistance to TKI treatment. The tripartite motif (TRIM) family member TRIM15 acts as an E3 ligase to mediate the polyubiquitination of substrates in cells. However, the biological role of TRIM15 in HCC is still an enigma. In our study, our results demonstrated that TRIM15 was abnormally upregulated in liver cancer cells after treated with TKIs and that this upregulation of TRIM15 contributed to TKI resistance in liver cancer cells. Then, we demonstrated that the upregulation of TRIM15 after TKI treatment was mediated by the AKT/FOXO1 axis. Moreover, we demonstrated that TRIM15 induced the nuclear translocation of LASP1 by mediating its K63-linked polyubiquitination, which modulated sensitivity to TKIs by increasing the phosphorylation of AKT and the expression of Snail in liver cancer cells. Collectively, we identified a novel AKT/FOXO1/TRIM15/LASP1 loop in cells, which provided potential candidates for overcoming TKI resistance in HCC.

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Section: Resultsmentioning

confidence: 99%

“…3d . In addition, we analyzed the RNA-seq data comparing the difference between regorafenib-resistant and -sensitive HCC cells reported by Sofer S et al [ 14 ]. (Supplementary Fig.…”

Section: Resultsmentioning

confidence: 99%

TRIM15 forms a regulatory loop with the AKT/FOXO1 axis and LASP1 to modulate the sensitivity of HCC cells to TKIs

et al. 2023

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“…Hexokinase (HK), the first glycolytic pathway enzyme, can be upregulated by hypoxia, which is associated with HCC resistance to regorafenib. 277 Mechanistically, hypoxia stabilizes HIF-1α by ubiquitin-specific peptidase (USP29) and then transcriptionally activates HK. 278 Pyruvate kinase M2 (PKM2), another rate-limiting enzyme of glycolysis, is overexpressed in TKI treatment, accompanied by high levels of lactate production.…”

Section: Mechanisms Of Tki Resistancementioning

confidence: 99%

Protein tyrosine kinase inhibitor resistance in malignant tumors: molecular mechanisms and future perspective

Yang

Wang

et al. 2022

Sig Transduct Target Ther

108

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Protein tyrosine kinases (PTKs) are a class of proteins with tyrosine kinase activity that phosphorylate tyrosine residues of critical molecules in signaling pathways. Their basal function is essential for maintaining normal cell growth and differentiation. However, aberrant activation of PTKs caused by various factors can deviate cell function from the expected trajectory to an abnormal growth state, leading to carcinogenesis. Inhibiting the aberrant PTK function could inhibit tumor growth. Therefore, tyrosine kinase inhibitors (TKIs), target-specific inhibitors of PTKs, have been used in treating malignant tumors and play a significant role in targeted therapy of cancer. Currently, drug resistance is the main reason for limiting TKIs efficacy of cancer. The increasing studies indicated that tumor microenvironment, cell death resistance, tumor metabolism, epigenetic modification and abnormal metabolism of TKIs were deeply involved in tumor development and TKI resistance, besides the abnormal activation of PTK-related signaling pathways involved in gene mutations. Accordingly, it is of great significance to study the underlying mechanisms of TKIs resistance and find solutions to reverse TKIs resistance for improving TKIs efficacy of cancer. Herein, we reviewed the drug resistance mechanisms of TKIs and the potential approaches to overcome TKI resistance, aiming to provide a theoretical basis for improving the efficacy of TKIs.

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“…Genome-scale perturbation with CRISPRa can be achieved by synthesizing a genome-wide sgRNA library, bypassing the need to generate a genome-wide ORF library. CRISPR-based GOF approaches have been used for genome-wide genetic screens in mammalian cells ( Konermann et al, 2015 ; Rodríguez et al, 2022 ; Zhu et al, 2022 ; Sofer et al, 2022 ; Zhang et al, 2022 ), but the feasibility has not yet been tested in Drosophila cells.…”

Section: Introductionmentioning

confidence: 99%

Pooled genome-wide CRISPR activation screening for rapamycin resistance genes in Drosophila cells

Xia

Viswanatha

et al. 2023

eLife

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Loss-of-function and gain-of-function genetic perturbations provide valuable insights into gene function. In Drosophila cells, while genome-wide loss-of-function screens have been extensively used to reveal mechanisms of a variety of biological processes, approaches for performing genome-wide gain-of-function screens are still lacking. Here, we describe a pooled CRISPR activation (CRISPRa) screening platform in Drosophila cells and apply this method to both focused and genome-wide screens to identify rapamycin resistance genes. The screens identified three genes as novel rapamycin resistance genes: a member of the SLC16 family of monocarboxylate transporters (CG8468), a member of the lipocalin protein family (CG5399), and a zinc finger C2H2 transcription factor (CG9932). Mechanistically, we demonstrate that CG5399 overexpression activates the RTK-Akt-mTOR signaling pathway and that activation of insulin receptor (InR) by CG5399 requires cholesterol and clathrin-coated pits at the cell membrane. This study establishes a novel platform for functional genetic studies in Drosophila cells.

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A genome‐wide CRISPR activation screen reveals Hexokinase 1 as a critical factor in promoting resistance to multi‐kinase inhibitors in hepatocellular carcinoma cells

Cited by 18 publications

References 41 publications

TRIM15 forms a regulatory loop with the AKT/FOXO1 axis and LASP1 to modulate the sensitivity of HCC cells to TKIs

TRIM15 forms a regulatory loop with the AKT/FOXO1 axis and LASP1 to modulate the sensitivity of HCC cells to TKIs

Protein tyrosine kinase inhibitor resistance in malignant tumors: molecular mechanisms and future perspective

Pooled genome-wide CRISPR activation screening for rapamycin resistance genes in Drosophila cells

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