Type II CRISPR/Cas9 approach in the oncological therapy

Biagioni, Alessio; Chillà, Anastasia; Andreucci, Elena; Laurenzana, Anna; Margheri, Francesca; Peppicelli, Silvia; Rosso, Mario Del; Fibbi, Gabriella

doi:10.1186/s13046-017-0550-0

Cited by 17 publications

(12 citation statements)

References 74 publications

(30 reference statements)

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“…RNA interference was performed as previously described [ 20 , 23 ]. The sequences targeting NUCB2–1 were: 5- CCACAGATTTAGATATGCTAA-3 and NUCB2–2 5-GCGTGAATATCATCAGGTCAT-3.…”

Section: Methodsmentioning

confidence: 99%

Regulation of the adaptation to ER stress by KLF4 facilitates melanoma cell metastasis via upregulating NUCB2 expression

Zhang

Lin

Chao

et al. 2018

J Exp Clin Cancer Res

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BackgroundAdaptation to ER stress has been indicated to play an important role in resistance to therapy in human melanoma. However, the relationship between adaptation to ER stress and cell metastasis in human melanoma remains unclear.MethodsThe relationship of adaptation to ER stress and cell metastasis was investigated using transwell and mouse metastasis assays. The potential molecular mechanism of KLF4 in regulating the adaptation to ER stress and cell metastasis was investigated using RNA sequencing analysis, q-RT-PCR and western blot assays. The transcriptional regulation of nucleobindin 2 (NUCB2) by KLF4 was identified using bioinformatic analysis, luciferase assay, and chromatin immunoprecipitation (ChIP). The clinical significance of KLF4 and NUCB2 was based on human tissue microarray (TMA) analysis.ResultsHere, we demonstrated that KLF4 was induced by ER stress in melanoma cells, and increased KLF4 inhibited cell apoptosis and promoted cell metastasis. Further mechanistic studies revealed that KLF4 directly bound to the promoter of NUCB2, facilitating its transcription. Additionally, an increase in KLF4 promoted melanoma ER stress resistance, tumour growth and cell metastasis by regulating NCUB2 expression in vitro and in vivo. Elevated KLF4 was found in human melanoma tissues, which was associated with NUCB2 expression.ConclusionOur data revealed that the promotion of ER stress resistance via the KLF4-NUCB2 axis is essential for melanoma cell metastasis, and KLF4 may be a promising specific target for melanoma therapy.Electronic supplementary materialThe online version of this article (10.1186/s13046-018-0842-z) contains supplementary material, which is available to authorized users.

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“…RNA interference was performed as previously described [ 20 , 23 ]. The sequences targeting NUCB2–1 were: 5- CCACAGATTTAGATATGCTAA-3 and NUCB2–2 5-GCGTGAATATCATCAGGTCAT-3.…”

Section: Methodsmentioning

confidence: 99%

Regulation of the adaptation to ER stress by KLF4 facilitates melanoma cell metastasis via upregulating NUCB2 expression

Zhang

Lin

Chao

et al. 2018

J Exp Clin Cancer Res

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“…As a follow-up of our studies, we have used the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)/Cas9 technique to obtain the irreversible clearing of uPAR in A375p and A375M6 human melanoma and in HCT116 colon cancer cell lines. This technology, based on a naturally-occurring system that protects bacteria from viral infections [17], is now widely used to manipulate genes in cells and living organisms [18][19][20]. Here we show that the uPAR KO clones have undergone a deep metabolic reprogramming, since the uPAR clearing in melanoma and colon carcinoma cell lines impairs the glycolytic pathway while enhancing the mitochondrial spare respiratory capacity and altering mitochondria biogenesis.…”

Section: Introductionmentioning

confidence: 83%

uPAR Knockout Results in a Deep Glycolytic and OXPHOS Reprogramming in Melanoma and Colon Carcinoma Cell Lines

Biagioni

Laurenzana

Chillà

et al. 2020

Cells

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Urokinase Plasminogen Activator (uPA) Receptor (uPAR) is a well-known GPI-anchored three-domain membrane protein with pro-tumor roles largely shown in all the malignant tumors where it is over-expressed. Here we have exploited the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 gene knock out approach to investigate its role in the oxidative metabolism in human melanoma and colon cancer as the consequences of its irreversible loss. Knocking out PLAUR, a uPAR-encoding gene, in A375p, A375M6 and HCT116, which are two human melanoma and a colon carcinoma, respectively, we have observed an increased number of mitochondria in the two melanoma cell lines, while we evidenced an immature biogenesis of mitochondria in the colon carcinoma culture. Such biological diversity is, however, reflected in a significant enhancement of the mitochondrial spare respiratory capacity, fueled by an increased expression of GLS2, and in a decreased glycolysis paired with an increased secretion of lactate by all uPAR KO cells. We speculated that this discrepancy might be explained by an impaired ratio between LDHA and LDHB.

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“…This complex system, which has been adapted to be used in laboratory practice, can recognize and cut DNA to provide a complete and high selective gene editing in vitro and in vivo. The possibility to be used in clinical treatment for several genetic derived pathologies has rapidly spread its fame worldwide [ 6 ]. Another obstacle in the race for the perfect gene therapy is that, the therapeutic translation of the CRISPR/Cas9 system lacks an appropriate delivery carrier [ 7 ].…”

Section: Introductionmentioning

confidence: 99%

Delivery systems of CRISPR/Cas9-based cancer gene therapy

et al. 2018

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CRISPR/Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats) is today one of the most reliable method for gene-editing, supporting previous gene therapies technologies such as TALEN, Meganucleases and ZFNs. There is a growing up number of manuscripts reporting several successful gene-edited cancer cell lines, but the real challenge is to translate this technique to the clinical practice. While treatments for diseases based on a single gene mutation is closer, being possible to target and repair the mutant allele in a selective way generating specific guide RNAs (gRNAs), many steps need to be done to apply CRISPR to face cancer. In this review, we want to give a general overview to the recent advancements in the delivery systems of the CRISPR/Cas9 machinery in cancer therapy.

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Type II CRISPR/Cas9 approach in the oncological therapy

Cited by 17 publications

References 74 publications

Regulation of the adaptation to ER stress by KLF4 facilitates melanoma cell metastasis via upregulating NUCB2 expression

Regulation of the adaptation to ER stress by KLF4 facilitates melanoma cell metastasis via upregulating NUCB2 expression

uPAR Knockout Results in a Deep Glycolytic and OXPHOS Reprogramming in Melanoma and Colon Carcinoma Cell Lines

Delivery systems of CRISPR/Cas9-based cancer gene therapy

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