Mass spectrometry for the study of adipocyte cell secretome in cardiovascular diseases

Gianazza, Erica; Brioschi, Maura; Eligini, Sonia; Banfi, Cristina

doi:10.1002/mas.21812

Cited by 5 publications

(2 citation statements)

References 125 publications

(176 reference statements)

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“…Mass spectrometry-based proteomic studies of adipocyte cell and tissue secretomes have identified hundreds of differentially expressed proteins (see Gianazza et al . [ 17 ] for a recent comprehensive review on the subject). The most relevant study for our work is on proteomic profiling of conditioned medium collected from primary human adipocytes [ 18 ].…”

Section: Fractionation Of Adipocyte-conditioned Medium Reveals Factor...mentioning

confidence: 99%

Native adiponectin plays a role in the adipocyte-mediated conversion of fibroblasts to myofibroblasts

El-Hattab

Sinclair

Liszewski

et al. 2023

J. R. Soc. Interface.

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Adipocytes regulate tissues through production of adipokines that can act both locally and systemically. Adipocytes also have been found to play a critical role in regulating the healing process. To better understand this role, we developed a three-dimensional human adipocyte spheroid system that has an adipokine profile similar to in vivo adipose tissues. Previously, we found that conditioned medium from these spheroids induces human dermal fibroblast conversion into highly contractile, collagen-producing myofibroblasts through a transforming growth factor beta-1 (TGF-β1) independent pathway. Here, we sought to identify how mature adipocytes signal to dermal fibroblasts through adipokines to induce myofibroblast conversion. By using molecular weight fractionation, heat inactivation and lipid depletion, we determined mature adipocytes secrete a factor that is 30–100 kDa, heat labile and lipid associated that induces myofibroblast conversion. We also show that the depletion of the adipokine adiponectin, which fits those physico-chemical parameters, eliminates the ability of adipocyte-conditioned media to induce fibroblast to myofibroblast conversion. Interestingly, native adiponectin secreted by cultured adipocytes consistently elicited a stronger level of α-smooth muscle actin expression than exogenously added adiponectin. Thus, adiponectin secreted by mature adipocytes induces fibroblast to myofibroblast conversion and may lead to a phenotype of myofibroblasts distinct from TGF-β1-induced myofibroblasts.

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Section: Fractionation Of Adipocyte-conditioned Medium Reveals Factor...mentioning

confidence: 99%

Native adiponectin plays a role in the adipocyte-mediated conversion of fibroblasts to myofibroblasts

El-Hattab

Sinclair

Liszewski

et al. 2023

J. R. Soc. Interface.

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“…date biofluid biomarkers for non-invasive cancer detection and disease monitoring [4][5][6]. To identify the cancer proteins secreted by cancer cells, mass spectrometry-based secretome proteomics has emerged as a powerful unbiased approach for marker identification [7][8][9][10][11][12].…”

Section: Introductionmentioning

confidence: 99%

Secretome processing for proteomics: A methods comparison

Almeida‐Marques,

Rolfs,

Piersma

et al. 2024

Proteomics

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The cancer cell secretome comprises a treasure‐trove for biomarkers since it reflects cross‐talk between tumor cells and their surrounding environment with high detectability in biofluids. In this study, we evaluated six secretome sample processing workflows coupled to single‐shot mass spectrometry: (1) Protein concentration by ultrafiltration with a molecular weight cut‐off (MWCO) filter and sample preparation through in‐gel digestion (IGD); (2) Acetone protein precipitation coupled to IGD; (3) MWCO filter‐based protein concentration followed by to in‐solution digestion (ISD); (4) Acetone protein precipitation coupled to ISD; (5) Direct ISD; (6) Secretome lyophilization and ISD. To this end, we assessed workflow triplicates in terms of total number of protein identifications, unique identifications, reproducibility of protein identification and quantification and detectability of small proteins with important functions in cancer biology such as cytokines, chemokines, and growth factors. Our findings revealed that acetone protein precipitation coupled to ISD outperformed the other methods in terms of the number of identified proteins (2246) and method reproducibility (correlation coefficient between replicates (r = 0.94, CV = 19%). Overall, especially small proteins such as those from the classes mentioned above were better identified using ISD workflows. Concluding, herein we report that secretome protein precipitation coupled to ISD is the method of choice for high‐throughput secretome proteomics via single shot nanoLC‐MS/MS.

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Fibroblast-Adipocyte Lineage Cell Interactions Result in Differential Production of Extracellular Matrix Proteins

Sander,

El-Hattab,

Jacobson

et al. 2024

Cel. Mol. Bioeng.

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Mass spectrometry for the study of adipocyte cell secretome in cardiovascular diseases

Cited by 5 publications

References 125 publications

Native adiponectin plays a role in the adipocyte-mediated conversion of fibroblasts to myofibroblasts

Native adiponectin plays a role in the adipocyte-mediated conversion of fibroblasts to myofibroblasts

Secretome processing for proteomics: A methods comparison

Fibroblast-Adipocyte Lineage Cell Interactions Result in Differential Production of Extracellular Matrix Proteins

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