TY - JOUR
T1 - Ultrasonic-assisted extraction and digestion of proteins from solid biopsies followed by peptide sequential extraction hyphenated to MALDI-based profiling holds the promise of distinguishing renal oncocytoma from chromophobe renal cell carcinoma
AU - Jorge, Susana
AU - Pereira, Kevin
AU - López-Fernández, Hugo
AU - LaFramboise, William
AU - Dhir, Rajiv
AU - Fernández-Lodeiro, Javier
AU - Lodeiro, Carlos
AU - Santos, Hugo M.
AU - Capelo-Martínez, Jose L.
N1 - PROTEOMASS Scientific Society is acknowledged by the funding provided to the Laboratory for Biological Mass Spectrometry Isabel Moura. Authors acknowledge the funding provided by the Associate Laboratory for Green Chemistry LAQV which is financed by national funds from FCT / MEC ( UID/QUI/50006/2019 ). H. M. S. is funded by the FCT 2015 Investigator Program ( IF/00007/2015 ). S. J. thanks FCT / MEC (Portugal) for her research contract as PhD student with the grant SFRH/BD/120537/2016 . J.F.L. Acknowledges FCT/MEC (Portugal) SFRH/BPD/93982/2013 and FCT-UNL for the DL57/2016 Assistant Researcher Contract. H. López-Fernández is supported by a post-doctoral fellowship from Xunta de Galicia ( ED481B 2016/068-0 ). This project utilized the University of Pittsburgh Hillman Cancer Center shared resource facility (Cancer Genomics Facility) supported in part by award P30CA047904 (Dr. LaFramboise).
PY - 2020/1/1
Y1 - 2020/1/1
N2 - A novel analytical approach is proposed to discriminate between solid biopsies of chromophobe renal cell carcinoma (chRCC) and renal oncocytoma (RO). The method comprises the following steps: (i) ultrasonic extraction of proteins from solid biopsies, (ii) protein depletion with acetonitrile, (iii) ultrasonic assisted in-solution digestion using magnetic nanoparticle with immobilized trypsin, (iv) C18 tip-based preconcentration of peptides, (v) sequential extraction of the peptides with ACN, (vi) MALDI-snapshot of the extracts and (vii) investigation of the extract containing the most discriminating features using high resolution mass spectrometry. With this approach we have been able to differentially cluster renal oncocytoma and chromophobe renal cell carcinoma and identified 18 proteins specific to chromophobe and seven unique to renal oncocytoma. Chromophobes express proteins associated with ATP function (ATP5I & 5E; VATE1 & G2; ADT2), glycolysis (PGK1) and neuromedin whilst oncocytomas express ATP5H, ATPA, DEPD7 and TRIPB thyroid receptor interacting protein.
AB - A novel analytical approach is proposed to discriminate between solid biopsies of chromophobe renal cell carcinoma (chRCC) and renal oncocytoma (RO). The method comprises the following steps: (i) ultrasonic extraction of proteins from solid biopsies, (ii) protein depletion with acetonitrile, (iii) ultrasonic assisted in-solution digestion using magnetic nanoparticle with immobilized trypsin, (iv) C18 tip-based preconcentration of peptides, (v) sequential extraction of the peptides with ACN, (vi) MALDI-snapshot of the extracts and (vii) investigation of the extract containing the most discriminating features using high resolution mass spectrometry. With this approach we have been able to differentially cluster renal oncocytoma and chromophobe renal cell carcinoma and identified 18 proteins specific to chromophobe and seven unique to renal oncocytoma. Chromophobes express proteins associated with ATP function (ATP5I & 5E; VATE1 & G2; ADT2), glycolysis (PGK1) and neuromedin whilst oncocytomas express ATP5H, ATPA, DEPD7 and TRIPB thyroid receptor interacting protein.
KW - Chromophobe renal cell carcinoma
KW - MALDI
KW - Profiling ESI
KW - Renal cancer
KW - Renal oncocytoma
KW - Sequential extraction
UR - http://www.scopus.com/inward/record.url?scp=85069894204&partnerID=8YFLogxK
U2 - 10.1016/j.talanta.2019.120180
DO - 10.1016/j.talanta.2019.120180
M3 - Article
C2 - 31514886
AN - SCOPUS:85069894204
SN - 0039-9140
VL - 206
JO - Talanta
JF - Talanta
M1 - 120180
ER -
