Oxidative Stress and Mitochondrial TERT in Papillary Thyroid Cancer.

Istituto Auxologico Italiano100 enrolled

Overview

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.

Study Type

OBSERVATIONAL

Enrollment

100

Conditions

Papillary Thyroid Cancer

Eligibility

Sex: ALL

Medical Language ↔ Plain English

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

Locations (1)

Outcomes

Primary Outcomes

H202 generation (nmol/mg tissue) in papillary thyroid cancer and in corresponding normal tissues.

Time frame: 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.

Time frame: 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.

Time frame: month 19-30

Mitochondrial oxidative stress generation in cells lines treated with Src kinase inhibitors.

Mitochondrial oxidative stress will be measured by immunofluorescence.

Time frame: months 25-36

Proliferation in cells lines treated with Src kinase inhibitors.

Proliferation will be measured by a colorimetric assay.

Time frame: months 25-36

Apoptosis in cells lines treated with Src kinase inhibitors.

Apoptosis will be measured by a luminometric assay.

Time frame: months 25-36

Migration in cells lines treated with Src kinase inhibitors.

Migration will be measured by wound healing assays.

Time frame: months 25-36

Secondary Outcomes

Genetic characterization of tumor tissues.

Point mutations and fusions will be investigated in DNA and RNA, respectively,

Time frame: 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.

Time frame: months 1-24