Immunohistochemistry and Next-Generation Sequencing for NTRK Fusion detection in Differentiated Thyroid Cancer of Children, Adolescents and Young Adults

Author:

Telles Ana Clara Tosta1,von Ammon Juliana Lima1,Matta Rafael Reis Campos1,Beltrão Fabyan Esberard de Lima2,Paz Alexandre Rolim2,Hecht Fábio3,Lopes Guilherme Castro4,Machado Gabriel Jeferson1,Cerqueira Taíse Lima de Oliveira1,Lisboa Bruno da Silva5,Rios Leonardo Freitas Boaventura5,Ramos Helton Estrela1

Affiliation:

1. Federal University of Bahia

2. Federal University of Paraíba

3. Federal University of Rio de Janeiro

4. Department of Molecular Genetics of Pardini Group

5. State University of Feira de Santana

Abstract

Abstract Introduction: Pan-Trk immunohistochemistry (IHC) staining can assess the protein expression from NTRK gene fusions a- Little is known about its utility in differentiated thyroid cancer (DTC) samples from children, adolescents, and young adults (CAYA) patients. Objective: Investigate Pan-Trk IHC sensitivity and specificity in DTC samples from CAYA patients. Methods:Tumor samples obtained from 79 CAYA patients (age <21years) diagnosed with DTC between January 2010 and January 2021 were retrospectively recruited from four health centers from state of Bahia e Paraíba, Brazil. NTRK gene fusion testing of all archival FFPE tumor samples: Pan-TRK IHC staining for TRKA, TRKB and TRKC protein expression were performed and then analyzed with RNA-based next-generation sequencing (NGS) assay in order to confirm IHC pan-TRK result and elucidate fusion partner. Results:Pan-Trk IHC: 3 of 79 cases had positive pan-Trk expression: NGS sequencing: 04- were identified with NTRK gene fusion, Pan-Trk IHC was negative in all 4 NTRK NGS-positive cases. 25 of 79 NTRK NGS-negative control cases had concordant negative pan-TRK IHC results. Therefore, our rate of false positive pan-Trk IHC results was 3/25 (12%). The overall results for pan-Trk IHC in our cohort of NGS-negative cases was: (i) sensitivity (0%), (ii) specificity (96%), (iii) positive predictive value (94.7%), (iv) negative predictive value (91%). Conclusions: Pan-Trk IHC was not a tissue-efficient screen for NTRK fusions in DTC from CAYA patients. This is the largest cohort of CAYA DTC cases stained with pan-Trk IHC, and it is the first to detail the sensitivity and specificity of pan-Trk IHC regarding the data obtained by targeted RNA-based NGS panel in DTC.

Publisher

Research Square Platform LLC

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