A putative OTU domain-containing protein 1 deubiquitinating enzyme is differentially expressed in thyroid cancer and identifies less-aggressive tumours

Abstract

Background:This study aimed to identify novel biomarkers for thyroid carcinoma diagnosis and prognosis.Methods:We have constructed a human single-chain variable fragment (scFv) antibody library that was selected against tumour thyroid cells using the BRASIL method (biopanning and rapid analysis of selective interactive ligands) and phage display technology.Results:One highly reactive clone, scFv-C1, with specific binding to papillary thyroid tumour proteins was confirmed by ELISA, which was further tested against a tissue microarray that comprised of 229 thyroid tissues, including: 110 carcinomas (38 papillary thyroid carcinomas (PTCs), 42 follicular carcinomas, 30 follicular variants of PTC), 18 normal thyroid tissues, 49 nodular goitres (NG) and 52 follicular adenomas. The scFv-C1 was able to distinguish carcinomas from benign lesions (P=0.0001) and reacted preferentially against T1 and T2 tumour stages (P=0.0108). We have further identified an OTU domain-containing protein 1, DUBA-7 deubiquitinating enzyme as the scFv-binding antigen using two-dimensional polyacrylamide gel electrophoresis and mass spectrometry.Conclusions:The strategy of screening and identifying a cell-surface-binding antibody against thyroid tissues was highly effective and resulted in a useful biomarker that recognises malignancy among thyroid nodules and may help identify lower-risk cases that can benefit from less-aggressive management.

Biotechnological and Immunological Platforms Based on PGL-I Carbohydrate-Like Peptide of Mycobacterium leprae for Antibodies Detection Among Leprosy Clinical Forms

“…The PGLI-M3 positivity for IgM was detected in MB clinical forms (BB, BL, and LL) follows similar patterns observed for the native PGL-I. Our study observed an average seropositivity of 78% in MB forms, while in the literature the positivity varied from 51.2 to 97.4% for the native PGL-I (Carneiro et al, 2014) and 76.36% for synthetic derivatives with ND-O-HSA ELISA (Lobato et al, 2011). The total positivity including all clinical forms was also similar, for example the lateral flow system (ML-Flow) using NT-P-HSA as antigen presented 97.4% (Bührer-Sékula et al, 2003), which corroborates our findings that showed 92.5% positivity using the PGLI-M3.…”

“…The selection of synthetic peptide PGLI-M3 binding antibodies was done using a scFv library fused to PIII protein (Carneiro et al, 2014) according to the protocol described elsewhere (Barbas, 2001). The combinatory scFv library, obtained from an RNA pool of non-infected individuals, with a 2 × 10 8 diversity, was amplified using a phage helper properly titrated.…”

Targeting the Ubiquitin Proteasome System in Cancer

“…Recently, OTUB1 is reported to be overexpressed in non-small-cell lung carcinoma (NSCLC) and promotes RAS activation by inhibiting RAS monoubiquitination [132]. Moreover, high expression of OTUD1 is also seen in thyroid carcinoma signifying its oncogenic nature [133]. On the other hand, OTUD5 is linked to apoptosis and is involved in stabilization and activation of p53, suggesting a possible tumor suppressive role [134].…”

Deubiquitinases and the new therapeutic opportunities offered to cancer

“…2009 ). The expression of OTUD1 is highly up-regulated in various carcinomas of the thyroid and could potentially be used as a biomarker to distinguish between cancerous samples ( Carneiro et al . 2014 ).…”