1 Scientific RepoRts | 7: 2649 | DOI:10.1038/s41598-017-02069-5 www.nature.com/scientificreports probing the action of a novel anti-leukaemic drug therapy at the single cell level using modern vibrational spectroscopy techniques Joanna L. Denbigh 1,2 , David perez-Guaita 3 , Robbin R. Vernooij 3 , Mark J. tobin 4 , Keith R. Bambery 4 , Yun Xu 1 , Andrew D. southam 5 , Farhat L. Khanim 5 , Mark T. Drayson 6 , Nicholas P. Lockyer 1 , Royston Goodacre 1 & Bayden R. Wood 3 Acute myeloid leukaemia (AML) is a life threatening cancer for which there is an urgent clinical need for novel therapeutic approaches. A redeployed drug combination of bezafbrate and medroxyprogesterone acetate (BaP) has shown anti-leukaemic activity in vitro and in vivo. elucidation of the BaP mechanism of action is required in order to understand how to maximise the clinical beneft. Attenuated total refectance Fourier transform infrared (ATR-FTIR) spectroscopy, Synchrotron radiation FTIR (S-FTIR) and Raman microspectroscopy are powerful complementary techniques which were employed to probe the biochemical composition of two AML cell lines in the presence and absence of BaP. Analysis was performed on single living cells along with dehydrated and fxed cells to provide a large and detailed data set. A consideration of the main spectral diferences in conjunction with multivariate statistical analysis reveals a signifcant change to the cellular lipid composition with drug treatment; furthermore, this response is not caused by cell apoptosis. No change to the DNA of either cell line was observed suggesting this combination therapy primarily targets lipid biosynthesis or efects bioactive lipids that activate specifc signalling pathways. Acute myeloid leukaemia (AML) is an aggressive cancer that leads to a build-up of non-diferentiated, dys- functional cells from the myeloid lineage. Without treatment, patients can die from impaired haemopoiesis within weeks of diagnosis and unfortunately responses to current treatments and overall survival of patients post-treatment generally remains poor 1 . Current AML treatment is centred on short cycles of intensive cytotoxic chemotherapy to kill the cancer cells, however, this treatment can disrupt the production of healthy blood cells. Continuous use of intensive chemotherapy is precluded because of the high levels of toxicity that afect haemopoiesis and cell division, which is important in maintaining the integrity of the body’s mucosal surfaces. Tis dose-limiting toxicity prevents escalation of chemotherapy to treat resistant disease and precludes its use in many older frailer individuals who account for a high proportion of AML patients 2 . Accordingly, there is an urgent need for low toxicity anti-AML therapies that can be used both as an adjunct to chemotherapy and in between chemotherapy cycles. Continuous low toxicity therapy is a novel strategy to manage AML by providing good quality survival even without total eradication of the disease. 1 Manchester Institute of Biotechnology and School of Chemistry, University of Manchester, Manchester, M1 7DN, United Kingdom. 2 Biomedical Research centre, School of environment and Life Sciences, University of Salford, Salford, M5 4WT, United Kingdom. 3 centre for Biospectroscopy and School of chemistry, Monash University, Clayton, Victoria, 3800, Australia. 4 Australian Synchrotron, 800 Blackburn Road, Clayton, Victoria, 3168, Australia. 5 School of Biosciences, University of Birmingham, Birmingham, B15 2TT, United Kingdom. 6 institute of immunology and Immunotherapy, University of Birmingham, Birmingham, B15 2TT, United Kingdom. Correspondence and requests for materials should be addressed to B.R.W. (email: Bayden.Wood@monash.edu) Received: 7 July 2016 Accepted: 7 February 2017 Published: xx xx xxxx OPEN