αCT1 peptide sensitizes glioma cells to temozolomide in a glioblastoma organoid platform

Che, Jingru; DePalma, Thomas; Sivakumar, Hemamylammal; Mezache, Louisa; Tallman, Miranda M.; Venere, Monica; Swindle-Reilly, Katelyn E; Veeraraghavan, Rengasayee; Skardal, Aleksander

doi:10.1002/bit.28313

Cited by 8 publications

(4 citation statements)

References 47 publications

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“…Few human cell lines exist. Future work in the ACC research field should be targeted towards determining the viability and clinical relevance of primary patient-derived ACC organoids—work that our team has moved into, that is an extension of our work with patient-derived tumor organoids in other cancers 17 , 34 , 36 , 64 , 68 – 70 . Such patient-derived tumor models can better retain the heterogeneity observed in tumors in vivo, and thus will serve as more appropriate systems in which to screen novel therapeutics.…”

Section: Discussionmentioning

confidence: 99%

A 3D adrenocortical carcinoma tumor platform for preclinical modeling of drug response and matrix metalloproteinase activity

Dedhia,

Sivakumar,

Rodriguez

et al. 2023

Sci Rep

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Adrenocortical carcinoma (ACC) has a poor prognosis, and no new drugs have been identified in decades. The absence of drug development can partly be attributed to a lack of preclinical models. Both animal models and 2D cell cultures of ACC fail to accurately mimic the disease, as animal physiology is inherently different than humans, and 2D cultures fail to represent the crucial 3D architecture. Organoids and other small 3D in vitro models of tissues or tumors can model certain complexities of human in vivo biology; however, this technology has largely yet to be applied to ACC. In this study, we describe the generation of 3D tumor constructs from an established ACC cell line, NCI-H295R. NCI-H295R cells were encapsulated to generate 3D ACC constructs. Tumor constructs were assessed for biomarker expression, viability, proliferation, and cortisol production. In addition, matrix metalloproteinase (MMP) functionality was assessed directly using fluorogenic MMP-sensitive biosensors and through infusion of NCI-H295R cells into a metastasis-on-a-chip microfluidic device platform. ACC tumor constructs showed expression of biomarkers associated with ACC, including SF-1, Melan A, and inhibin α. Treatment of ACC tumor constructs with chemotherapeutics demonstrated decreased drug sensitivity compared to 2D cell culture. Since most tumor cells migrate through tissue using MMPs to break down extracellular matrix, we validated the utility of ACC tumor constructs by integrating fluorogenic MMP-sensitive peptide biosensors within the tumor constructs. Lastly, in our metastasis-on-a-chip device, NCI-H295R cells successfully engrafted in a downstream lung cell line-based construct, but invasion distance into the lung construct was decreased by MMP inhibition. These studies, which would not be possible using 2D cell cultures, demonstrated that NCI-H295R cells secreted active MMPs that are used for invasion in 3D. This work represents the first evidence of a 3D tumor constructs platform for ACC that can be deployed for future mechanistic studies as well as development of new targets for intervention and therapies.

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Section: Discussionmentioning

confidence: 99%

A 3D adrenocortical carcinoma tumor platform for preclinical modeling of drug response and matrix metalloproteinase activity

Dedhia,

Sivakumar,

Rodriguez

et al. 2023

Sci Rep

Self Cite

View full text Add to dashboard Cite

show abstract

“…Future work in the ACC research field should be targeted towards determining the viability and clinical relevance of primary patient-derived ACC organoids -work that our team has moved into, that is an extension of our work with patient-derived tumor organoids in other cancers. 17,34,36,64,[68][69][70] Such patient-derived tumor models can better retain the heterogeneity observed in tumors in vivo, and thus will serve as more appropriate systems in which to screen novel therapeutics. Our MOC platform to date has primarily been used to model metastasis kinetics, largely without any therapeutic intervention.…”

Section: Discussionmentioning

confidence: 99%

A 3D adrenocortical carcinoma tumor platform for preclinical modeling of drug response and matrix metalloproteinase activity

Dedhia

Sivakumar

Rodríguez

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

Adrenocortical carcinoma (ACC) has a poor prognosis, and no new drugs have been identified in decades. The absence of drug development can partly be attributed to a lack of preclinical models. Both animal models and 2D cell cultures of ACC fail to accurately mimic the disease, as animal physiology is inherently different than humans, and 2D cultures fail to represent the crucial 3D architecture. Organoids are small 3D in vitro models of tissues or tumors, which can model certain complexities of human in vivo biology; however, this technology has yet to be applied to ACC. In this study, we describe the generation of organoids from an established ACC cell line, H295R. H295R cells were encapsulated to generate ACC organoid constructs. Organoids were assessed for biomarker expression, viability, proliferation, and cortisol production. In addition, matrix metalloproteinase (MMP) functionality was assessed directly using fluorogenic MMP-sensitive biosensors and through infusion of H295R cells into a metastasis-on-a-chip microfluidic device platform. ACC organoids showed expression of biomarkers associated with ACC, including SF-1, Melan A, and inhibin alpha. Treatment of ACC organoids with chemotherapeutics demonstrated decreased drug sensitivity compared to 2D cell culture. Since most tumor cells migrate through tissue using MMPs to break down extracellular matrix, we validated the utility of ACC organoids by integrating fluorogenic MMP-sensitive peptide biosensors within the organoids. Lastly, in our metastasis-on-a-chip device, H295R cells successfully engrafted in a downstream lung cell line-based organoid construct, but invasion into the lung organoid construct was decreased by MMP inhibition. These studies, which would not be possible using 2D cell cultures, demonstrated that H295R cells secreted active MMPs that are used for migration in 3D. This work represents the first evidence of a 3D organoid platform for ACC that can be deployed for future mechanistic studies as well as development of new targets for intervention and therapies.

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“…Glioma cells overexpressing Cx43 exhibit higher resistance to TMZ, while Cx43 knockdown, in the same model system, sensitizes these cells to TMZ, implying a crucial role for this connexin in modulating chemoresistance in GBM. These ndings suggest that combining Cx43 inhibitors with TMZ could offer a valid therapeutic approach for TMZ-resistant GBM patients (15,16,19,(22)(23)(24). Finally, Cx43 overexpression in tumor cells promotes migration and invasion, and its expression level positively correlates with invasive capacity in GBM cell lines (25)(26)(27).…”

Section: Introductionmentioning

confidence: 91%

Chlorpromazine Overcomes Temozolomide Resistance in Glioblastoma by inhibiting Cx43 and Essential DNA Repair Pathways

Matarrese,

Signore,

Ascione

et al. 2024

Preprint

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Background In the fight against GBM, drug repurposing emerges as a viable and time-saving approach to explore new treatment options. Chlorpromazine, an old antipsychotic medication, has recently arisen as a promising candidate for repositioning in GBM therapy in addition to temozolomide, the first-line standard of care. We previously demonstrated the antitumor efficacy of chlorpromazine and its synergistic effects with temozolomide in suppressing GBM cell malignant features in vitro. This prompted us to accomplish a Phase II clinical trial to evaluate the efficacy and safety of adding chlorpromazine to temozolomide in GBM patients with unmethylated MGMT gene promoter. In this in vitro study, we investigate the potential role of chlorpromazine in overcoming temozolomide resistance. Methods In our experimental set, we analyzed Connexin-43 expression at both the transcriptional and protein levels in control- and chlorpromazine-treated GBM cells. DNA damage and subsequent repair were assessed by immunofluorescence of g-H2AX and Reverse-Phase Protein microArrays in chlorpromazine treated GBM cell lines. To elucidate the relationship between DNA repair systems and chemoresistance, we analyzed a signature of DNA repair genes in GBM cells after treatment with chlorpromazine, temozolomide and Connexin-43 downregulation. Results Chlorpromazine treatment significantly downregulated connexin-43 expression in GBM cells, consequently compromising connexin-dependent cellular resilience, and ultimately contributing to cell death. In line with this, we observed concordant post-translational modifications of molecular determinants involved in DNA damage and repair pathways. Our evaluation of DNA repair genes revealed that temozolomide elicited an increase, while chlorpromazine, as well as connexin-43 silencing, a decrease in DNA repair gene expression in GBM cells. Conclusions Chlorpromazine potentiates the cytotoxic effects of the alkylating agent temozolomide through a mechanism involving downregulation of Cx43 expression and disruption of the cell cycle arrest essential for DNA repair processes. This finding suggests that chlorpromazine may be a potential therapeutic strategy to overcome TMZ resistance in GBM cells by inhibiting their DNA repair mechanisms.

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αCT1 peptide sensitizes glioma cells to temozolomide in a glioblastoma organoid platform

Cited by 8 publications

References 47 publications

A 3D adrenocortical carcinoma tumor platform for preclinical modeling of drug response and matrix metalloproteinase activity

A 3D adrenocortical carcinoma tumor platform for preclinical modeling of drug response and matrix metalloproteinase activity

A 3D adrenocortical carcinoma tumor platform for preclinical modeling of drug response and matrix metalloproteinase activity

Chlorpromazine Overcomes Temozolomide Resistance in Glioblastoma by inhibiting Cx43 and Essential DNA Repair Pathways

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