Elucidating the role of cardiolipin in mitochondrial and cellular functions

Current funding for this project:

NIH – R01HL117880-05 – The Role of Cardiolipin in the TCA Cycle: Implications for Barth Syndrome

Barth Syndrome Foundation – Supplementation of critical metabolites improves TCA cycle function and viability of tafazzin-deficient cells

AHA Predoctoral Fellowship (to Zhuqing Liang) – Frataxin deficiency in cardiolipin-deficient cells leads to defective Fe-S biogenesis

Cardiolipin (CL), the signature phospholipid of mitochondria, is crucial not only for mitochondrial function but also for a plethora of cellular processes that are not associated with mitochondrial bioenergetics. The de novo synthesis of CL is followed by a remodeling cycle of deacylation (removal of a fatty acid) followed by reacylation. Specific fatty acyl composition is acquired during this process, and remodeled CL contains predominantly unsaturated fatty acids.  The importance of CL remodeling is underscored by the life-threatening genetic disorder Barth syndrome (BTHS), caused by mutations in tafazzin, the enzyme that reacylates CL.

CL-deficient mutants have been instrumental in elucidating the functions of this unique lipid.  We identified the first CL mutant (crd1Δ), which cannot synthesize CL. We also developed the yeast model for BTHS, the taz1Δ mutant, and a mouse myoblast BTHS model, TAZ-KO, which lack tafazzin. Utilizing these mutants, we have determined that CL is required for optimal mitochondrial bioenergetics and energy metabolism; mitochondrial protein import; mitochondrial fusion; function of the vacuole, the vacuolar ATPase, and vacuolar autophagy; iron-sulfur biogenesis; and MAPK signaling. Furthermore we have shown that genetically blocking the CL deacylation step rescues the defective phenotypes of the tafazzin mutant.

Our current studies are focused on elucidating the mechanisms whereby CL controls these diverse cellular activities.  Our approach is to utilize the power of the yeast model to develop hypotheses that can be tested in appropriate mammalian systems. Specifically, our research addresses the following questions:

  1. How does CL deficiency perturb cellular energy metabolism?
  2. What is the mechanism linking CL to iron-sulfur biogenesis?
  3. What is the role of CL in MAPK signaling, vacuolar function, and autophagy?
  4. What is the function of CL remodeling?
  5. What defects associated with CL deficiency lead to the pathologies in BTHS and other CL-associated disorders, including diabetic cardiomyopathy, heart failure, and ischemia/reperfusion injury?


Cellular consequences of inositol depletion

Current funding for this project:

NIH – R01GM125082-07 – Novel Mechanisms of Regulation of Inositol Biosynthesis

Inositol is an essential metabolite that plays a fundamental role in regulating cellular processes. The phosphorylation of inositol generates numerous inositol phosphates and phosphoinositides, many of which are signaling molecules that control membrane trafficking, calcium mobilization, chemotaxis, ion channel activity, cytoskeletal organization, and gene expression. Furthermore, phosphatidylinositol is a precursor for the synthesis of sphingolipids, which play an important role in signal transduction and trafficking.  Consistent with an essential role in cellular function, starvation for inositol leads to a rapid loss of viability. The crucial role of inositol-containing compounds in cell function is underscored by the link between perturbation of inositol metabolism and human disorders, including bipolar disorder, Alzheimer’s and Parkinson’s diseases, malignant hyperthermia, cancer, type 2 diabetes, Lowe syndrome, myotubular myopathy, and Charcot-Marie-Tooth disease. Therefore, elucidating the mechanisms underlying the control of inositol homeostasis is expected to have important implications for a broad range of illnesses.

The long-term goal of our research is to elucidate the molecular mechanisms underlying regulation of inositol homeostasis, to identify the cellular consequences of inositol depletion, and to determine the role of inositol depletion in the therapeutic mechanisms of drugs used to treat bipolar disorder.  To this end, we have identified three novel mechanisms of regulation of inositol synthesis – inhibition of synthesis by phosphorylation of the rate-limiting biosynthetic enzyme myo-inositol-3-P synthase (MIPS), perturbation of MIPS transcription by inositol pyrophosphates, and modulation of inositol synthesis by glycogen kinase 3 (GSK3).  In addition, we have shown that the drug valproate (VPA) causes inositol depletion in yeast and human cells by indirectly inhibiting MIPS.  We have determined that a novel consequence of inositol depletion is perturbation of the vacuolar ATPase (V-ATPase) by altering homeostasis of the phosphoinositide PI3,5P2.  The V-ATPase is required for acidification and function of intracellular compartments, including lysosomes, secretory vesicles and synaptic vesicles. The proton gradient generated by the V-ATPase is utilized in the uploading and storage of neurotransmitters.  Therefore, our studies suggest that the therapeutic effects of inositol depletion by valproate may be mediated in part by perturbation of the V-ATPase.Using the powerful yeast genetic model to elucidate the molecular mechanisms that control inositol homeostasis and mammalian cells to test the hypotheses generated from the yeast model, we are addressing the following questions:

  1. What are the molecular mechanisms underlying regulation of inositol synthesis by phosphorylation of MIPS, pyrophosphate regulation of MIPS transcription, and GSK3-activation?
  2. What is the mechanism whereby inositol-depleting drugs inhibit synthesis of inositol in yeast and human cells?
  3. How does inositol depletion perturb the V-ATPase?
  4. Do inositol-depleting drugs affect sphingolipid metabolism?
  5. How do inositol-depleting drugs perturb neurotransmission?