Oxidative Stress and Mitochondrial TERT in Papillary Thyroid Cancer.

Study Overview
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Brief Summary
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Oxidative stress (OS) could be involved in the progression of papillary thyroid cancer (PTC). Indeed, thyroid differentiation genes are silenced by a mechanism controlled by NOX4-derived OS. On the other hand, TERT contributes to mitochondrial OS protection, which could increase the resistance of cancer cells to therapeutic agents. The investigators aim to address the role of OS and mitochondrial TERT in the progression and therapeutic resistance of PTC. OS and TERT subcellular localization will be investigated in 150 PTCs and correlated to the genetic and expression profile of the tumors and to the clinical and prognostic features of the patients. Mechanisms implicated in TERT mitochondrial migration and the contribution of mitochondrial TERT to tumor progression will be investigated in cancer cell lines and primary cell cultures. This study will allow to identify OS as a marker of therapeutic resistance in PTC and will open new opportunities for the development of novel treatments targeting ROS generation/TERT nuclear export.

Official Title
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The Role of Oxidative Stress and Mitochondrial TERT in the Progression and Therapeutic Resistance of Papillary Thyroid Cancer.

Conditions
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Papillary Thyroid Cancer

Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Patients with papillary thyroid cancer Patients whose tumor and contralateral healthy tissues can be collected for the analyses Exclusion Criteria: patients who did not provide consent patients lost at follow-up tissues not adequate for the analyses
Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| H202 generation (nmol/mg tissue) in papillary thyroid cancer and in corresponding normal tissues. |  | months 1-24 | 
| TERT mitochondrial localization (TERT/VDAC) in papillary thyroid tumors. | TERT mitochondrial localization will be investigated by Western blot of mitochondrial fractions and normalized to VDAC protein expression. | months 13-30 | 
| Effect of exogenous oxidative stress (H2O2) and therapeutic agents (BRAF, MEK and Src kinase inhibitors) on TERT nuclear to mitochondrial translocation in thyroid cancer cell lines. | TERT mitochondrial translocation will be measured by immunofluorescence. | month 19-30 | 
| Mitochondrial oxidative stress generation in cells lines treated with Src kinase inhibitors. | Mitochondrial oxidative stress will be measured by immunofluorescence. | months 25-36 | 
| Proliferation in cells lines treated with Src kinase inhibitors. | Proliferation will be measured by a colorimetric assay. | months 25-36 | 
| Apoptosis in cells lines treated with Src kinase inhibitors. | Apoptosis will be measured by a luminometric assay. | months 25-36 | 
| Migration in cells lines treated with Src kinase inhibitors. | Migration will be measured by wound healing assays. | months 25-36 | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Genetic characterization of tumor tissues. | Point mutations and fusions will be investigated in DNA and RNA, respectively, | months 1-24 | 
| Expression profile of tumor tissues. | The expression level of 16 thyroid function genes will be investigated by a custom RNA sequencing panel. | months 1-24 |