serum lipidomic study reveals potential early biomarkers for predicting response to chemoradiotherapy in advanced rectal cancer a pilot study

Accepted Manuscript Serum Lipidomic study reveals potential early biomarkers for predicting response to chemoradiotherapy in advanced rectal cancer: a pilot study Piero Del Boccio, Francesca Perrotti, Claudia Rossi, Ilaria Cicalini, Sara Di Santo, Mirco Zucchelli, Paolo Sacchetta, Domenico Genovesi, Damiana Pieragostino PII: S2452-1094(16)30103-8 DOI: 10.1016/j.adro.2016.12.005 Reference: ADRO 59 To appear in: Advances in Radiation Oncology Please cite this article as: Del Boccio P, Perrotti F, Rossi C, Cicalini I, Di Santo S, Zucchelli M, Sacchetta P, Genovesi D, Pieragostino D, Serum Lipidomic study reveals potential early biomarkers for predicting response to chemoradiotherapy in advanced rectal cancer: a pilot study, Advances in Radiation Oncology (2017), doi: 10.1016/j.adro.2016.12.005 This is a PDF file of an unedited manuscript that has been accepted for publication As a service to our customers we are providing this early version of the manuscript The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain ACCEPTED MANUSCRIPT Serum Lipidomic study reveals potential early biomarkers for predicting response to chemoradiotherapy in advanced rectal cancer: a pilot study Del BoccioPiero1,2 , Perrotti Francesca3,4, Rossi Claudia 2,5, Cicalini Ilaria 1,2, Di Santo Sara 3,4, Zucchelli Mirco2, Sacchetta Paolo2,5, Genovesi Domenico 3,4 and Pieragostino Damiana 2,5 * Department of Pharmacy, University “G d’Annunzio” of Chieti-Pescara, Chieti, Italy Analitical Biochemistry and Proteomics Unit, Research Centre on Aging (Ce.S.I), University “G d’Annunzio” of Chieti-Pescara, Chieti, Italy RI PT Department of Neurosciences and Imaging, University “G d’Annunzio” of Chieti-Pescara, SC Chieti, Italy Radiation Oncology Unit, SS Annunziata Hospital in Chieti – Italy Department of Medical Oral and Biotechnological Sciences, University “G d’Annunzio” of Chieti-Pescara, Chieti, Italy TE D *Corresponding Author: M AN U Dr DamianaPieragostino, PhD Department of Medical Oral and Biotechnological Sciences, University “G d’Annunzio” Chieti-Pescara, Chieti, Italy EP e-mail: dpieragostino@unich.it Phone: +39 0871 541593 AC C Fax: +39 0871 541598 Conflict of interest statements Dr Pieragostino has nothing to disclose Prof Del BoccioPiero has nothing to disclose Dr Perrotti Francesca has nothing to disclose Dr Rossi Claudia has nothing to disclose Dr Cicalini Ilaria has nothing to disclose Dr Di Santo Sara has nothing to disclose ACCEPTED MANUSCRIPT Dr Zucchelli Mirco has nothing to disclose Prof Sacchetta Paolo has nothing to disclose AC C EP TE D M AN U SC RI PT Prof Genovesi Domenico has nothing to disclose ACCEPTED MANUSCRIPT Serum Lipidomic study reveals potential early biomarkers for predicting response to chemoradiotherapy in advanced rectal cancer: a pilot study Keywords: Lipidomics; Chemoradiotherapy; Biomarkers; Personalized Medicine; Advanced Rectal RI PT Cancer Summary This study aims to highlight a typical lipid signature able to predict the tumor response to preoperative chemoradiotherapy in advanced rectal cancer, by using a Lipidomics approach SC Five lipids were validated as biomarkers able to predict response before treatment, resulting in a ROC curve characterized by an (area under the curve) AUC of 0.95 Results suggest as M AN U serum lipids could represent an useful tool in prediction of CRT response, towards a personalized treatment Introduction Colorectal Cancer (CRC) is the third most frequently cancer globally[1] Preoperative fluoropyrimidine-based chemoradiotherapy (CRT) or short-course radiotherapy (RT) TE D followed by total mesorectal excision (TME) are the standard treatments for CRC [2-4] To date, there is an increasing interest in predicting which patients will respond to neoadjuvant CRT[5], in the effort to personalize treatments, especially in investigating easy accessible EP biological fluids [6], and to improve response rate and survival outcomes Several biomarkers have been investigated for their ability to predict outcome in LARC treated with CRT, but few works investigated lipids[7-9] Bioactive lipids are fundamental mediators of a number AC C of biological processes[10-12] and the implication of lipids in cancer growth and diffusion was already demonstrated[13] In this work we aimed to study serum polar lipid in a prospective cohort of LARC patients before CRT (t0 group), including patients naïve to chemotherapy and radiotherapy Samples were also collected during CRT (t14 and t28 days), in the effort to correlate the global lipid signature to response to treatment Methods (see supplementary materials) ACCEPTED MANUSCRIPT Results Lipidomics biomarker discovery Serum from 18 patients with LARC (7 females and 11 males), of which classified as Responder (RP) and 10 subjects as Not Responder (NRP) according to Mandard’s tumor RI PT regression grading (TRG) and treated with preoperative CRT was analyzed by LC-ESIMS/MS Data were converted into a matrix containing m/z signals coupled with retention time as variables and the patient codes as observations This dataset was reduced by considering only variables present in at least 50% of patients Figure shows the lipid classes (including lyso forms) screened in the four panels The studied lipids were SC Sphingomyelins (SMs) and Phosphatidylcholines (PCs) in panel A, Phatidylethanolamines (PEs) in panel B, Phosphatidylglycerols (PGs) in Panel C and Phosphatidylserines (PS) in M AN U Panel D Each lipid class screened, was reported in the four panels In particular in Figure panel A the score plot of PC/SM phospholipids is shown, while panel B shows the score plot of PE class; in panel C the PG lipids are shown and the PS class is reported in panel D The resulting PLS-DA models are reported as score scatter plots in Figure 1, showing clear separation between RP and NRP before treatment The lipids identified as Variable Important for the Projection (VIP>1) were confirmed through univariate test At t0, 65 lipids were TE D identified as significant with the criteria of VIP>1.5 and p