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Label-free quantitative data dependent (DDA) nano-LC-MS/MS proteomic profiling of mouse lymph harvested from mesenteric and cervical anatomical districts

ABSTRACT: To fully quantify differences in the lymph composition and associated dendritic cells antigenic load, from different anatomical districts and in physiological and pathological conditions, we collected the afferent lymph draining to the cervical and mesenteric lymph nodes in healthy mice. Most of the proteome present in the mesenteric afferent lymph, showed a profile of proteins involved in different metabolic pathways associated with lipoproteintransport, and lipid metabolism, such as adipocyte-type fatty acid binding protein, Apolipoproteins A, B, C and E, and phospholipid transfer proteins, consistent with the known role of the mesenteric lymph in chylomicron transport. Network analysis on the mesenteric afferent lymph unique/enriched proteome highlighted pathways associated with lipase and hydrolase activity, lipoproteins remodeling, fat digestion and absorption, triglyceride catabolism and gut-associated immune cells and cytokines responses. Among the proteome shared across tissue a brain-specific or highly enriched proteome including glia maturation factor, nerve growth factor, mesencephalic astrocyte-derived neurotrophic factor, alpha-crystallin, brain-specific isoform of glycogen phosphorylase, and proteins associated with voltage-dependent channels, were uniquely observed in the lymph harvested from the afferent lymphatics entering the deep cervical nodes. Network analysis on the afferent cervical unique/enriched proteome highlighted pathways associated with neurotransmitter release cycle, synaptic transmission, neuronal development, mitochondrial activity, and an overall CNS proteome.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Mus Musculus (ncbitaxon:10090)

SUBMITTER: Cristina C Clement Laura Santambrogio

PROVIDER: MSV000094586 | MassIVE | Sun Apr 21 17:21:00 BST 2024

SECONDARY ACCESSION(S): PXD051618

REPOSITORIES: MassIVE

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Project description:Purpose: Presence of pelvic lymph node metastases is the main prognostic factor in early stage cervical cancer patients, primarily treated with surgery. Aim of this study was to identify cellular tumor pathways associated with pelvic lymph node metastasis in early stage cervical cancer. Experimental Design: Gene expression profiles (Affymetrix U133 plus 2.0) of 20 patients with negative (N0) and 19 with positive lymph nodes (N+), were compared with gene sets that represent all 285 presently available pathway signatures. Validation immunostaining of tumors of 274 consecutive early stage cervical cancer patients was performed for representatives of the identified pathways. Results: Analysis of 285 pathways resulted in identification of five pathways (TGF-β, NFAT, ALK, BAD, and PAR1) that were dysregulated in the N0, and two pathways (β-catenin and Glycosphingolipid Biosynthesis Neo Lactoseries) in the N+ group. Class comparison analysis revealed that five of 149 genes that were most significantly differentially expressed between N0 and N+ tumors (P<0.001) were involved in β-catenin signaling (TCF4, CTNNAL1, CTNND1/p120, DKK3 and WNT5a). Immunohistochemical validation of two well-known cellular tumor pathways (TGF-β and β-catenin) confirmed that the TGF-β pathway (positivity of Smad4) was related to N0 (OR:0.20, 95%CI:0.06-0.66) and the β-catenin pathway (p120 positivity) to N+ (OR:1.79, 95%CI:1.05-3.05). Conclusions: Our study provides new, validated insights in the molecular mechanism of lymph node metastasis in cervical cancer. Pathway analysis of the microarray expression profile suggested that the TGF-β and p120-associated non-canonical β-catenin pathways are important in pelvic lymph node metastasis in early stage cervical cancer. For the microarray experiment, we selected fresh frozen primary cervical cancer tissue, containing at least 80% tumor cells, of patients with histologically confirmed N0 (n=20) and of patients with N+ (n=19). The N0 and N+ groups were matched for age, FIGO stage and histology (all squamous cell carcinoma).

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Label-free quantitative data dependent (DDA) nano-LC-MS/MS proteomic profiling of mouse lymph harvested from mesenteric and cervical anatomical districts

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