Development of small-molecule inhibitors targeting adipose triglyceride lipase

Mayer, Nicole; Schweiger, Martina; Romauch, Matthias; Grabner, G; Eichmann, Thomas O.; Fuchs, Elisabeth; Ivkovic, Jakov; Heier, Christoph; Mrak, Irina; Lass, Achim; Höfler, Gerald; Fledelius, Christian; Zechner, Rudolf; Zimmermann, R.; Breinbauer, Rolf

doi:10.1038/nchembio.1359

Cited by 168 publications

(196 citation statements)

References 25 publications

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“…Based on our results, the most parsimonious explanation for the abnormal lipid peak observed by cerebral magnetic resonance spectroscopy in human subjects with DDHD2 mutations (6) is that it reflects elevated TAGs, although it remains possible that DDHD2 could regulate distinct classes of lipids in the mouse and human brain. We also cannot explain, at present, why subchronic treatment with a DDHD2 inhibitor is needed to produce TAG elevations in the mouse brain, but we should note that acute treatment with an ATGL inhibitor also causes more limited changes in TAGs in peripheral tissues compared with chronic (genetic) disruption of ATGL (22,33). Projecting forward, it will be important to evaluate how deregulated TAG metabolism and LD accumulation affect neuronal cell biology and, ultimately, HSP neuropathology.…”

Section: Discussionmentioning

confidence: 99%

The hereditary spastic paraplegia-related enzyme DDHD2 is a principal brain triglyceride lipase

Inloes

Hsu

Dix

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

143

184

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Complex hereditary spastic paraplegia (HSP) is a genetic disorder that causes lower limb spasticity and weakness and intellectual disability. Deleterious mutations in the poorly characterized serine hydrolase DDHD2 are a causative basis for recessive complex HSP. DDHD2 exhibits phospholipase activity in vitro, but its endogenous substrates and biochemical functions remain unknown. Here, we report the development of DDHD2 −/− mice and a selective, in vivo-active DDHD2 inhibitor and their use in combination with mass spectrometry-based lipidomics to discover that DDHD2 regulates brain triglycerides (triacylglycerols, or TAGs). DDHD2 −/− mice show age-dependent TAG elevations in the central nervous system, but not in several peripheral tissues. Large lipid droplets accumulated in DDHD2 −/− brains and were localized primarily to the intracellular compartments of neurons. These metabolic changes were accompanied by impairments in motor and cognitive function. Recombinant DDHD2 displays TAG hydrolase activity, and TAGs accumulated in the brains of wild-type mice treated subchronically with a selective DDHD2 inhibitor. These findings, taken together, indicate that the central nervous system possesses a specialized pathway for metabolizing TAGs, disruption of which leads to massive lipid accumulation in neurons and complex HSP syndrome. D etermining the genetic basis for rare hereditary human diseases has benefited from advances in DNA sequencing technologies (1). As a greater number of disease-causing mutations are mapped, however, it is also becoming apparent that many of the affected genes code for poorly characterized proteins. Assigning biochemical and cellular functions to these proteins is critical to achieve a deeper mechanistic understanding of human genetic disorders and for identifying potential treatment strategies.Hereditary spastic paraplegia (HSP) is a genetically heterogeneous neurologic syndrome marked by spasticity and lower extremity weakness (2). Many genetic types of HSP have been identified and are numbered according to their order of discovery [spastic paraplegia (SPG) 1-72] (2, 3). Of these genetic variants, more than 40 have been mapped to causative mutations in protein-coding genes. HSP genes code for a wide range of proteins that do not conform to a single sequence-or function-related class. A subset of HSP genes, including PNPLA6 (or neuropathytarget esterase) (SPG39) (4), DDHD1 (SPG28) (5), and DDHD2 (SPG54) (3, 6-8), code for serine hydrolases. These enzymes have been designated as (lyso)phospholipases based on in vitro substrate assays (9-11), but their endogenous substrates and physiological functions remain poorly understood. The mutational landscape that affects these lipid hydrolases to cause recessive HSP is complex but collectively represents a mix of null and putatively null and/or functional mutations. Moreover, the type of HSP appears to differ in each case, with DDHD1 mutations causing uncomplicated HSP, whereas PNPLA6 and DDHD2 mutations lead to complex forms of the disease that...

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Section: Discussionmentioning

confidence: 99%

The hereditary spastic paraplegia-related enzyme DDHD2 is a principal brain triglyceride lipase

Inloes

Hsu

Dix

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

143

184

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“…The Atgl / mice also provided a model to study the specificity of the ATGL-specific inhibitor, Atglistatin, in HSCs (22). Surprisingly, Atglistatin treatment for 6 days resulted in an almost similar increase in TAG levels in Atgl / HSCs as compared with WT HSCs (Fig.…”

Section: Atglistatin Inhibits the Predominant Lipase In Mouse Hscsmentioning

confidence: 97%

“…3). This would eliminate monoglyceride lipase, HSL, lipoprotein lipase, and pancreatic lipase, as well as PNPLA6 and PNPLA7, which were shown to be unaffected by this drug (22). PNPLA3, also called adiponutrin (ADPN), is also an unlikely candidate because this enzyme is involved in TAG remodeling, rather than in net lipolysis (26).…”

Section: Discussionmentioning

confidence: 99%

ATGL and DGAT1 are involved in the turnover of newly synthesized triacylglycerols in hepatic stellate cells

Tuohetahuntila

Molenaar

Spee

et al. 2016

Journal of Lipid Research

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“…A recent study on serine hydrolase inhibitors ( 18 ) using rodent tissues showed that WWL64 is a specifi c inhibitor of patatin-like phospholipase domain containing 2 /ATGL. Another study ( 17 ) reported ( 20 ), and also hCES1 ( Tables 1 and 2 ). The widely used DAGL inhibitor, RHC20867, actually barely inhibited hDAGL ␣ at 5 M but inhibited hHSL, hMAGL, hABHD6, and hCES1 to different degrees ( Table 1 ).…”

Section: Abhd6 Activity and Inhibitionmentioning

confidence: 99%

Simplified assays of lipolysis enzymes for drug discovery and specificity assessment of known inhibitors

Iglesias

Lamontagne

Erb

et al. 2016

Journal of Lipid Research

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is also an important source of several metabolic signals. As an integral part of the glycerolipid/fatty acid cycle ( 1 ), lipolysis produces lipid signals that modify cellular functions such as glucose-stimulated insulin secretion ( 2 ) and metabolic pathways and also alter transcription of various genes ( 1, 3 ). TG breakdown to glycerol and fatty acids is accomplished by the sequential action of adipose triglyceride lipase (ATGL), which hydrolyzes TG to 2,3-or 1,3-diacylglycerol (DAG), followed by hormone sensitive lipase (HSL)-mediated DAG hydrolysis to generate 1-or 2-monoacylglycerol (MAG) ( 1, 3 ). Finally MAG is hydrolyzed by either the classical MAG lipase (MAGL) or the recently described ␣ / ␤ -hydrolase domain 6 (ABHD6) to glycerol and FFA ( 4-6 ). Receptor-mediated signaling at the plasma membrane leads to the phospholipase-C-dependent formation of 1,2-DAG, which is further hydrolyzed by sn -1-DAG lipases (DAGL) ␣ or ␤ ( 7 ), to form mostly 2-MAG.Recent studies indicated the physiological importance of many of these lipases. Thus, ATGL has been implicated in lipid homeostasis in adipocytes, myocardium and skeletal muscle, cancer cachexia ( 1,3,8 ), and the regulation of insulin secretion in pancreatic ␤ -cells ( 9 ). HSL was shown to be important in adipose lipid metabolism ( 10 ) and in the regulation of glucose-stimulated insulin secretion ( 11,12 ). MAGL, which hydrolyzes the endocannabinoid Abstract Lipids are used as cellular building blocks and condensed energy stores and also act as signaling molecules. The glycerolipid/ fatty acid cycle, encompassing lipolysis and lipogenesis, generates many lipid signals. Reliable procedures are not available for measuring activities of several lipolytic enzymes for the purposes of drug screening, and this resulted in questionable selectivity of various known lipase inhibitors. We now describe simple assays for lipolytic enzymes, including adipose triglyceride lipase (ATGL), hormone sensitive lipase (HSL), sn -1-diacylglycerol lipase (DAGL), monoacylglycerol lipase, ␣ / ␤ -hydrolase domain 6, and carboxylesterase 1 (CES1) using recombinant human and mouse enzymes either in cell extracts or using purifi ed enzymes. We observed that many of the reported inhibitors lack specifi city. Thus, Cay10499 (HSL inhibitor) and RHC20867 (DAGL inhibitor) also inhibit other lipases. Marked differences in the inhibitor sensitivities of human ATGL and HSL compared with the corresponding mouse enzymes was noticed. Thus, ATGListatin inhibited mouse ATGL but not human ATGL, and the HSL inhibitors WWL11 and Compound 13f were effective against mouse enzyme but much less potent against human enzyme. Many of these lipase inhibitors also inhibited human CES1. Results describe reliable assays for measuring lipase activities that are amenable for drug screening and also caution about the specifi city of the many earlier described lipase inhibitors.

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Development of small-molecule inhibitors targeting adipose triglyceride lipase

Cited by 168 publications

References 25 publications

The hereditary spastic paraplegia-related enzyme DDHD2 is a principal brain triglyceride lipase

The hereditary spastic paraplegia-related enzyme DDHD2 is a principal brain triglyceride lipase

ATGL and DGAT1 are involved in the turnover of newly synthesized triacylglycerols in hepatic stellate cells

Simplified assays of lipolysis enzymes for drug discovery and specificity assessment of known inhibitors

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