Vim 3 antibodyUse of Vimentin 3 for the diagnosis and differentiation of benign and malignant renal carcinoma Prof. Dr. Jochen Fries, Dr. Melanie von Brandenstein Facts about kidney cancer • ca. 338,000 new cases worldwide each year (World Cancer Research Fund) • kidney tumors: benign or malignant • malignant tumors: 3 subtypes according to their morphological appearance • 1. Clear cell renal cell carcinoma (RCC) (81%) 2. Papillary renal cell carcinoma (10%) 3. Chromophobe renal cell carcinoma (4%) • benign tumor: Oncocytoma (5%) The importance of correct classification Renal Benign Carcinoma Oncocytoma Operation Renal vein infiltration Lymph node (LN) resection Follow - up Kidney removal >Tu-enucleation about 20% frequently necessary if LN + if tumor in vein + Metastases >10yrs ++ (bone, lung,adrenal) Death ++ Tu-enucleation > Kidney removal none none none none none Current standard diagnosis for tumor differentiation • Immune histology: Malignant RCC: Combination of antibodies against cytokeratins AND Vimentin • RCC subtypes positive for Vimentin (full length) • Oncocytoma should be Vimentin (full length) negative Correct diagnosis determines operative procedure Clear cell Papillary Chromophobe Oncocytic Why do we need an antibody to identify the Oncocytoma? 1) Operative planning, follow-up, and survival risk assessment is the same for any subtype of malignant RCC but different in Oncocytoma 2) No antibody specific for any renal tumor subtype 3) Malignant RCC subtypes have eosinophilic variants, which mimic Oncocytoma !! 4) Vimentin (marker for malignant RCC) is positive in 20% of all Oncocytomas ( false positivity??) 5) Tumor regression greatly compromises light microscopic diagnosis Problem: Renal tumors are often regressive; Difficulties when biopsied Malignant RCC eosinophilic subtypes can mimic Oncocytoma Benign oncocytoma?? Renal cell carcinoma?? SOLUTION !?! Vimentin 3 Antibody • • • • What is it? Specificity and sensitivity Applicability Usefulness in daily diagnostics Solution - the new HOPE – Vim 3 V3 Antikörper The Antibody 3B4 Antikörper SP20 Antikörper Vimentin FL ~ 57 kDa V9 Antikörper Vimentin 3 ~ 47 kDa Solution – Vim 3 antibody Established on paraffinized tissue and by Western blot proof of specificity via blast against the designed AA sequence: only Vim 3 with this kDa size only Vim 3 has the exact AA sequence AA of the C-terminal ending of Vim 3 is 8 AA Solution – tumor collective Design of specific primers which only detect the Vim 3 variant proof of specificity via Sanger Sequenzing of the PCR product qRT-PCR results of the tumor collective qRT-PCR results of Vim 3 expression in the different tumor types -ß-actin was used as normalization Immune histology Full length Vim3 Immune flourescence Vim3 Vimentin full length Malignant RCC eosinophilic subtypes can mimic Oncocytoma Benign oncocytoma?? Renal cell carcinoma?? Eosinophilic Tumor = Papillary Carcinoma FL V3 Benign Oncocytoma FL FL V3 V3 Conclusion • Vimentin3 (Vim3) antibody detects only the Vim 3 isoform of Vimentin (high specificity) = Differentiation marker of Vimentin positivity • Vim 3 identifies specifically benign oncocytoma : a) First specific antibody for renal tumors b) Renal cell carcinoma and all subtypes are negative for Vim 3 expression • Convenient and easy detection by routine methods, e. g. immune histology, immune fluorescence • Applicable for Western blotting (high sensitivity) • Handling requires only normal pathology lab routine IP-Status - Developmental Status - Offer IP-Status: PCT application has been filed Developmental Status: 20 tumors (fresh tissue; own archive) 40 tumors (paraffinized tissue;Prof.Störkel, Wuppertal) qRT-PCR (triplicates); Matching immune histology/fluorescence (duplicates) results } Offer: In-licensing and co-development contact: PROvendis GmbH, Kordula Kruber Thank you for your attention! K idney O NCOCYTOMA E valuation L aboratory V imentiN3
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