DataSheet1_Metabolomic profiling of triple negative breast cancer cells suggests that valproic acid can enhance the anticancer effect of cisplatin.docx

Cisplatin is an effective chemotherapeutic agent for treating triple negative breast cancer (TNBC). Nevertheless, cisplatin-resistance might develop during the course of treatment, allegedly by metabolic reprograming, which might influence epigenetic regulation. We hypothesized that the histone deacetylase inhibitor (HDACi) valproic acid (VPA) can counter the cisplatin-induced metabolic changes leading to its resistance. We performed targeted metabolomic and real time PCR analyses on MDA-MB-231 TNBC cells treated with cisplatin, VPA or their combination. 22 (88%) out of the 25 metabolites most significantly modified by the treatments, were acylcarnitines (AC) and three (12%) were phosphatidylcholines (PCs). The most discernible effects were up-modulation of AC by cisplatin and, contrarily, their down-modulation by VPA, which was partial in the VPA-cisplatin combination. Furthermore, the VPA-cisplatin combination increased PCs, sphingomyelins (SM) and hexose levels, as compared to the other treatments. These changes predicted modulation of different metabolic pathways, notably fatty acid degradation, by VPA. Lastly, we also show that the VPA-cisplatin combination increased mRNA levels of the fatty acid oxidation (FAO) promoting enzymes acyl-CoA synthetase long chain family member 1 (ACSL1) and decreased mRNA levels of fatty acid synthase (FASN), which is the rate limiting enzyme of long-chain fatty acid synthesis. In conclusion, VPA supplementation altered lipid metabolism, especially fatty acid oxidation and lipid synthesis, in cisplatin-treated MDA-MB-231 TNBC cells. This metabolic reprogramming might reduce cisplatin resistance. This finding may lead to the discovery of new therapeutic targets, which might reduce side effects and counter drug tolerance in TNBC patients.</p

Additional file 1: of Safety and efficacy of human embryonic stem cell-derived astrocytes following intrathecal transplantation in SOD1G93A and NSG animal models

Figure S1. hES-AS produce and secrete neurotrophic factors. Conditioned media of 24 h from cultures of hES-AS differentiated for 28 days as well as cell extracts used to measure level of neurotrophic factors GDNF, BDNF, VEGF and IGF-1. For each factor, bars show cell content, amount secreted and negative control (medium only), expressed in pg/106 cells (triplicates ¹ SD) (PDF 91 kb

Additional file 4: of Safety and efficacy of human embryonic stem cell-derived astrocytes following intrathecal transplantation in SOD1G93A and NSG animal models

Table S2. Percent of cell presence and percent of frequency scores greater than, or equal to ‘2’ (one to three foci of 10-20 cells per foci) for each follow up time (4, 17 and 39 weeks after hES-AS transplantation). Supplementary materials and methods. (ZIP 150 kb

Additional file 3: of Safety and efficacy of human embryonic stem cell-derived astrocytes following intrathecal transplantation in SOD1G93A and NSG animal models

Figure S2. Effect of hES-AS transplantation on disease onset, progression and survival in hSOD1G93A mice. hES-AS, differentiated for 7 days, transplanted intrathecally through CM of hSOD1G93A mice. A Three experimental groups tested, single injection of 2 × 106 hES-AS on day 67 of life (Cellsx1), two injections of 2 × 106 hES-AS each on days 67 and 97 (Cellsx2) and once sham-injected mice (vehicle). Kaplan–Meir plot of disease onset (measured by 3% body weight loss from maximal weight) showing more delay in twice-injected group. B Kaplan–Meier survival curves with similar trends. C Body weight maintained longer in hES-AS-treated mice. Note that a few days after second injection, day 97, weight loss occurred related to injection. D Neurological score. E Significant improvement in motor performance (Rotarod test) for hSOD1 mice transplanted twice with hES-AS. C, D Values are mean ± SEM (PDF 262 kb