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A disease of metabolism: Defining lipedema’s molecular hallmarks by multi-omics approach for disease prediction

ABSTRACT: Characterized by pathologic subcutaneous adipose tissue development in the limbs, lipedema is a chronic disease in women. Molecular disease pathways and potential drug target genes remain to be found. In this study, we investigated the molecular and cellular hallmarks of lipedema using a multi-omics approach. We found adipokine dysregulation in early stage lipedema patients, which points towards the adipocyte as a key player. Unexpectedly, lipedema patients’ serum lactosyl- and hexosyl-sphingolipids increased compared to control subjects, when adjusted for age. Additionally, we observed immune cell exclusion from adipose tissue in lipedema patients. In subjects with lipedema, both expression of genes in VEGF signaling pathways (blood vessel/vascular development) as well as VEGF protein levels in serum increased. Expression of genes in the inflammatory response pathway points towards the absence of adipose tissue inflammation, and is in line with reduced serum inflammatory markers IL5 and FLT3L. The systemic reduction of serum glutamic acid levels, regulation of protein and RNA glutamic—pyruvic transaminase and reduced glutathione levels could link reduced inflammation with metabolic processes. The protein pathways for blood coagulation, and regulation of body fluid levels are most strongly down regulated. There is a case to be made that lipedema could be caused by dysregulation of metabolism that leads to immune cell exclusion. These findings provide new insights into the pathogenesis of lipedema and suggest potential targets for therapeutic intervention. Last but not least we used machine learning to train support vector machine based models with either serum sphingolipids and adipokines or adipose tissue RNA to predict the lipedema phenotype at a high accuracy.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Adipose Tissue

SUBMITTER: Paul Langlais

LAB HEAD: Leon Straub

PROVIDER: PXD058489 | Pride | 2025-03-17

REPOSITORIES: Pride

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			Action	DRS
	20191114_C3P0_4298_1.raw	Raw
	20191114_C3P0_4298_1.raw__F006045_.mzid.gz	Mzid
	20191114_C3P0_4298_1.raw__F006045_.mzid_20191114_C3P0_4298_1.raw__F006045_.MGF	Mzid
	20191114_C3P0_4298_17.raw	Raw
	20191114_C3P0_4298_17.raw__F006189_.mzid.gz	Mzid

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Project description:During the colonization of hosts, bacterial pathogens are presented with many challenges that must be overcome for colonization to successfully occur. This requires bacterial sensing of the surroundings and adaptation to the conditions encountered. One of the major impediments to pathogen colonization of the mammalian gastrointestinal tract is the antibacterial action of bile. Salmonella enterica serovar Typhimurium has specific mechanisms involved in resistance to bile. Besides being resistant to it, Salmonella can also successfully multiply in bile, using it as a source of nutrients. This accomplishment is highly relevant to pathogenesis, as Salmonella colonizes the gallbladder of hosts, where it can be carried asymptomatically and promote further host spread and transmission. In order to gain insights into the mechanisms used by Salmonella to grow in bile, we studied the changes elicited by Salmonella in the chemical composition of bile during growth in vitro and in vivo through a metabolomics approach. Our data suggest that phospholipids are an important source of carbon and energy for Salmonella during growth in the laboratory as well as during gallbladder infections of mice. Further studies in this area will generate a better understanding of how Salmonella exploits this generally hostile environment for its own benefit. For in vitro studies, bile was extracted from C57BL/6 mice and used immediately. An overnight culture of Salmonella enterica serovar Typhimurium SL1344 was used to inoculate 10 ?L of bile at an approximate density of 5x10^6 cells/mL. The experiment was performed in duplicate, and a total of 2 uninfected and 2 infected bile samples were studied. Samples were incubated for 24 hours at 37 oC with shaking. After incubation, samples were centrifuged to remove bacteria and the supernatant was saved for metabolomic analyses. For in vivo studies, C57BL/6 mice were infected with approximately 10^8 bacterial cells by oral gavage. Four groups of three to four mice each were either infected with Salmonella or kept uninfected (total of 11 mice per treatment group). Five days after infection, all mice were sacrificed and bile was collected. Samples were centrifuged and supernatants saved for metabolomic analyses. Samples were prepared by mixing equal volumes of bile from three to four mice, generating three samples per treatment (uninfected and infected), which were used in the subsequent steps. Seven microliters of each sample was evaporated and the residue resuspended in 50% acetonitrine. Extracts were infused into a 12-T Apex-Qe hybrid quadrupole-FT-ICR mass spectrometer equipped with an Apollo II electrospray ionization source, a quadrupole mass filter and a hexapole collision cell. Raw mass spectrometry data were processed as described elsewhere (Han et al. 2008. Metabolomics. 4:128-140). To identify differences in metabolite composition between different groups of samples, we filtered the list of masses for metabolites which were present on one set of samples but not the other. Additionally, we calculated the ratios between averaged intensities of metabolites from each group of mice. To assign possible metabolite identities to the masses selected as described above, the monoisotopic neutral masses of interest were queried against the Human Metabolome Database (HMDB, http://www.hmdb.ca), with a tolerance of 0.001 Da.

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A disease of metabolism: Defining lipedema’s molecular hallmarks by multi-omics approach for disease prediction

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