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HIGH CD36 gene signature generated from upregulated genes in AML patients that express the most CD36 mRNA compared to that of the lowest expression of CD36 in TCGA cohort.

Supplementary Methods. Supplementary Figure S1. Clinical distributions of AML patients from TUH and 3 independent patient cohorts. Supplementary Figure S2. In vivo treatment with 60 mg/kg/d of cytarabine (AraC) given daily for 5 days induces a significant reduction of the total cell tumor burden in AML-engrafted mice. Supplementary Figure S3. In vivo cytarabine (AraC) treatment induces significant but heterogeneous response in bone marrow and spleen of AML-xenografted NSG mice. Supplementary Figure S4. Comparative analysis of the in vivo response to cytarabine (AraC) with clinicobiological data of AML patients. Supplementary Figure S5. In vivo cytarabine (AraC) treatment induces changes in CD34+/-CD38+/- phenotypes in AML-engrafted mice. Supplementary Figure S6. In vivo cytarabine (AraC) chemotherapy treatment leads to reduction of the absolute number of human CD34+CD38+/- populations in AML. Supplementary figure S7. No enrichment in G0 quiescent cells was observed in mice treated with sublethal dose of cytarabine (AraC) for 5 days in vivo. Supplementary figure S8. Mitochondrial and energetic features of LOW (KG1, KG1a, U937) and HIGH (MOLM14, MV4-11, HL60) OXPHOS AML cell lines. Supplementary figure S9. Functional analysis of the transcriptomes of LOW (KG1a, U937) versus HIGH (MOLM14, HL60) OXPHOS AML cell lines untreated or treated with metformin. Supplementary figure S10. AML cells surviving after cytarabine (AraC) treatment are resistant to chemotherapies and are pre-existing CD36+CD44+ phenotype with an increased oxidative metabolism. Supplementary figure S11. Culture in galactose induces energetic shift of LOW OXPHOS AML U937 cells toward HIGH OXPHOS state, leading to cytarabine (AraC) resistance in AML. Supplementary figure S12. Energetic shift of mtDNA-depleted Rho0 MOLM14 cells toward LOW OXPHOS state induces AraC sensitivity. Supplementary figure S13. Electron Transfer Chain Complex I inhibition by Phenformin (Phenf) induces energetic shift toward LOW OXPHOS state and increases cytarabine (AraC) sensitivity in MOLM14 cells. Supplementary figure S14. Electron Transfer Chain Complex I inhibition by Metformin (Met) induces energetic shift toward LOW OXPHOS state and increases cytarabine (AraC) sensitivity in MOLM14 cells. Supplementary figure S15. Electron Transfer Chain Complex I inhibition by Rotenone (Rot) induces energetic shift toward LOW OXPHOS state and increases cytarabine (AraC) sensitivity in MOLM14 cells. Supplementary figure S16. Electron Transfer Chain Complex III inhibition by Antimycin A (AntiA) induces energetic shift toward LOW OXPHOS state and increases cytarabine (AraC) sensitivity in MOLM14 cells. Supplementary figure S17. Electron Transfer Chain Complex III inhibition by Atovaquone (ATQ) induces energetic shift toward LOW OXPHOS state and increases cytarabine (AraC) sensitivity in MOLM14 cells. Supplementary figure S18. Working model of the resistance to AraC in vivo.

Analytical biochemical and molecular genetic studies, based on biopsies of muscle, skin and tissue culture have become standard methods for a more precise determination of mitochondrial defects. It is hoped that from these studies some precise classification of mitochondrial diseases will arise in the future. At present the information is incomplete due to a series of problems faced by mitochondrial researchers, not accustomed to deal with the precise requirements of human biopsy analytical methodology and with the lack of precise association between genetic and biochemical data. There is plenty of information, about 45,000 publications on mitochondrial structure and function and 2600 on mitochondrial diseases. However, we are in need of scientists that can combine the rigor of the clinical analytical biochemist with the specialized approach and inquisitiveness of the mitochondrial researcher. Therefore, discussion of methodological problems and standardization is especially important. Dr. Ernster provided an historical introduction and perspective of the exponential growth of information on mitochondrial diseases. This field grew from a single paper in 1959 to 2500 in 1996, and that now encompasses more than one hundred diseases including aging and chronic adult neurodegenerative diseases. He also discussed present work carried in his lab with CoQ deficiencies, presenting the pathways for the synthesis of CoQ and points where defects in the pathway can arise. He also discussed the vicious circle of mitochondrial DNA mutations and oxidative stress. Dr. C.P. Lee developed the important point, sometimes not easy to achieve, that only with isolated, coupled mitochondria, is it possible to start diagnosing mitochondrial defects with a certain degree of precision. If not done in such a way, confusion will result when attempting to correlate genotype with the biochemical lesion. She offered a precise example of a case where multiple defects in a patient were found only after exploring coupled mitochondria function not only with the oxygen electrode but also with spectral and stability studies of the components of the respiratory chain. Dr. Lee referred also to the risks of studying enzyme kinetics in homogenates, where side reactions can lead to erroneous results. Studies with homogenates requires the judicious use and titration with respiratory chain inhibitors to verify the absence of side reactions. Use of more than one assay method for the same enzymatic process is necessary to make sure that results are coherent. Dr. Trijbels discussed substrate transport defects, all nuclear genome-dependent effects. They cannot be studied unless isolated coupled mitochondria are used. Dr. Kunz presented microtechniques for the study of mitochondral function using single muscle fibers and introduced the use of computer imaging and fluorescence sorting as tools for diagnosis. While still highly specialized, his techniques offer an interesting and needed miniaturization approach to analytical procedures.