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Contaminants and detergents were removed using SP3-based precipitation and washing on 5 µL magnetic beads1. Briefly, 100 µL ethanol for precipitation, 50 µL ethanol for washing followed by drying of proteins in the speedvac before adding trypsin and Lys-C proteases. A second overnight digestion step was added to improve digestion efficiency as previously described2. Evotip PURE clean-up of human ganglion samples 1 µg of each tissue digest was desalted per Evotip. The Evotip PURE protocol was adjusted for offline C18 clean-up in a 96-well format as described previously3. Initially, Evotip PURE tips were rinsed with 20 µl of Buffer B (comprising 80% ACN, water and 0.1% formic acid) and spun down at 800g for 60 s. The Evotips were conditioned with 10 µl of isopropanol, followed by an 1-min spin at 100g and additional 1 min at 400g to empty the Evotips. The PURE Evotips were washed and equilibrated in 200 µl of Buffer A (0.1% formic acid). Samples were acidified in 5% TFA, and the Evotip PURE was emptied by centrifuging at 800g for 1 min. The acidified samples were loaded onto the PURE Evotips and spun at 800g for 1 min. The samples were washed with 200 µl of Buffer A and spun down at 800g for 1 min. Elutions were collected in PCR strips by eluting with 20 µl of Buffer B by centrifuging at 100g for 1 min followed by 450g 1 min. The peptides were dried in a SpeedVac and resuspended in 40ul of 0.1% TFA supplemented with 0.015% DDM for mass spectrometry analyses. Up to 2ul (or 50ng peptides) were injected per MS anlaysis. Liquid chromatography and mass spectrometry (LC-MS/MS) The mass spectrometry data for mouse samples was acquired in data-independent acquisition (DIA) mode. The LC-MS/MS analysis was carried out using EASY nanoLC 1200 (Thermo Fisher Scientific) coupled with trapped ion mobility spectrometry quadrupole time-of-flight single cell proteomics mass spectrometer (timsTOF SCP, Bruker Daltonik GmbH, Germany) via a CaptiveSpray nano-electrospray ion source. Peptides (50 ng) were loaded onto a 25 cm Aurora Series UHPLC column with CaptiveSpray insert (75 μm ID, 1.6 μm C18) at 50°C and separated using a 50 min gradient (5-20% buffer B in 30 min, 20-29% buffer B in 9 min, 29-45% in 6 min, 45-95% in 5 min, wash with 95% buffer B for 5 min, 95-5% buffer B in 5 min) at a flow rate of 300 nL/min. Buffer A and B were water with 0.1 vol% formic acid and 80:20:0.1 vol% ACN:water: formic acid, respectively. MS data were acquired in single-shot library- free DIA mode and the timsTOF SCP was operated in DIA/parallel accumulation serial fragmentation (PASEF) using the high sensitivity detection-low sample amount mode. The ion accumulation and ramp time were set to 100 ms each to achieve nearly 100% duty cycle. The collision energy was ramped linearly as a function of the mobility from 59 eV at 1/K0 = 1.6 Vs cm−2 to 20 eV at 1/K0 = 0.6 Vs cm−2. The isolation windows were defined as 24 X 25 Th from m/z 400 to 1000. The mass spectrometry data for human samples were acquired using the same modification as in SCP-nano, ie using 5.5 cm long PAC HT column to minimize carryover and to accelerate column cleaning and maintenance between different sample types3. References 1. Hughes, C. S. et al. Single-pot, solid-phase-enhanced sample preparation for proteomics experiments. Nat. Protoc. 14, 68–85 (2019). 2. Bhatia, H. S. et al. Spatial proteomics in three-dimensional intact specimens. Cell 185, 5040–5058 (2022). 3. Luo, J. et al. Nanocarrier imaging at single-cell resolution across entire mouse bodies with deep learning. Nat. Biotechnol. https://doi.org/10.1038/s41587-024-02528-1 (2025)."],"repository":["Pride"],"quantification_method":[""],"modification":[""],"data_protocol":["Proteomics data processing For mouse data, diaPASEF raw files were searched against the mouse uniport database using DIA-NN4. Peptides length range from seven amino acids were considered for the search including N-terminal acetylation. Oxidation of methionine was set as a variable modification and cysteine carbamidomethylation as fixed modification. Enzyme specificity was set to Trypsin/P with 2 missed cleavages. The FASTA digest for library-free search was enabled for predicting the library generation. The FDR was set to 1% at precursor and global protein level. Match-between-runs (MBR) feature was enabled and quantification mode was set to “Robust LC (high precision)”. The Protein Group column in DIA-NN’s report was used to identify the protein group and PG.MaxLFQ was used to calculate the differential expression. For human data, analogous searches using DIANN 2.0 were performed. Proteomics data analysis Mouse data were analyzed using scanpy (v. 1.10.1) and anndata (v. 0.8.0) in Python 3.10. Twelve independent samples were analyzed from each group (High-Fat Diet and Chow) from three animals with samples from both right and left trigeminal ganglia. All proteins expressed in less than half of the samples in each group were filtered out, resulting in 6686 proteins used for downstream analyses. The data was log-transformed and normalized per sample. The missing values were input using KNNImputer (n_neighbors=5) from sklearn package (v. 1.2.1). With scanpy’s dendrogram function scipy’s hierarchical linkage clustering was calculated on a Pearson correlation matrix over groups which was calculated for 50 averaged principal components.To identify differentially regulated proteins across two groups (HFD and Chow), we combined samples from the right and the left trigeminal ganglia. Differential expression analysis was conducted using Scanpy’s method ` rank_genes_groups` with method set to `t-test We applied a threshold of p < 0.05 and |log fold change| > 0.5 to identify differentially expressed proteins (DEPs). These DEPs were subsequently visualized using volcano plots. Pathway enrichment analysis was performed on the combined up- and down-regulated proteins using the KEGG and Reactome databases. The most relevant pathways were highlighted, displaying the DEPs involved in each pathway. Human data were analyzed analogously using the Fragpipe Analyst tool5 focusing on differentially abundant proteins and differentially regulated pathways using the human KEGG and reactome database resources.  References 1. Hughes, C. S. et al. Single-pot, solid-phase-enhanced sample preparation for proteomics experiments. Nat. Protoc. 14, 68–85 (2019). 2. Bhatia, H. S. et al. Spatial proteomics in three-dimensional intact specimens. Cell 185, 5040–5058 (2022). 3. Luo, J. et al. Nanocarrier imaging at single-cell resolution across entire mouse bodies with deep learning. Nat. Biotechnol. https://doi.org/10.1038/s41587-024-02528-1 (2025). 4. Demichev, V. et al. DIA-NN: neural networks and interference correction enable deep proteome coverage in high throughput. Nat. Methods 17, 41–44 (2020). 5. Hsiao, Y. et al. Analysis and Visualization of Quantitative Proteomics Data Using FragPipe-Analyst. J. Proteome Res. 23, 4303–4315 (2024)."],"omics_type":["Proteomics"],"labhead":["Ali Erturk"],"instrument_platform":[""],"submission_type":["PARTIAL"],"labhead_affiliation":["nstitute for Intelligent Biotechnologies, Helmholtz Center Munich, German Research Center for Environmental Health, Neuherberg, Germany Institute for Stroke and Dementia Research, Klinikum der Universität München, Ludwig-Maximilians-Universität LMU, Munich, German School of Medicine, Koç University, İstanbul, Turkey Deep Piction, Munich, Germany"],"species":["Homo Sapiens (human)"],"publication":["Not available"],"submitter_mail":["ali.erturk@helmholtz-munich.de"],"submitter_affiliation":["Institute for Tissue Engineering and Regenerative Medicine, Helmholtz Munich"],"submitter_country":["Germany"],"additional_accession":[]},"is_claimable":false,"name":"Deep Learning and 3D Imaging Reveal Whole-body Alterations in Obesity","description":"Many diseases, such as obesity, have systemic effects that perturb multiple organ systems throughout the body. However, tools for comprehensive, high-resolution analysis of disease-associated changes at the whole-body scale have been lacking. Here, we developed MouseMapper, a suite of foundation model-based deep learning algorithms, to enable a multi-system analysis of disease across the entire mouse body. MouseMapper enables quantitative, graph-based analysis of nerves and immune cells at the whole-body level resolving fine axonal branches and immune cell clusters and automatically segments 31 organs and tissues in the mouse body.  We applied MouseMapper to study high-fat diet-induced obesity, uncovering a significant degeneration of the infraorbital branch of the trigeminal ganglia. This structural damage in infraorbital nerves was associated with functional sensory deficits in whisker sensing. Furthermore, we identified proteomic changes in the trigeminal ganglion affecting axon remodeling and complement pathways both in mouse and human. MouseMapper also generated detailed 3D inflammation maps by characterizing immune cell cluster compositions across tissues. The MouseMapper framework demonstrates robust generalizability across different imaging resolutions and datasets. Our study provides a powerful, scalable approach for discovering and quantifying systemic pathologies, bridging molecular insights from animal models to human conditions.","dates":{"publication":"2026-05-06","submission":"2025-11-25"},"accession":"PXD071172","cross_references":{"TAXONOMY":["NEWT:108018","NEWT:377960","NEWT:1131","NEWT:190486","NEWT:2737160","NEWT:2042546","NEWT:148942","NEWT:193528","NEWT:295546","NEWT:258594","NEWT:1126","NEWT:1129","NEWT:89187","NEWT:309807","NEWT:89184","NEWT:309800","NEWT:57577","NEWT:156563","NEWT:1323429","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:2315688","NEWT:44270","NEWT:592321","NEWT:1073882","NEWT:6819","NEWT:93217","NEWT:428386","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:32264","NEWT:10034","NEWT:10031","NEWT:82569","NEWT:661410","NEWT:488003","NEWT:224326","NEWT:376619","NEWT:3765","NEWT:2433","NEWT:1190602","NEWT:3767","NEWT:433727","NEWT:135622","NEWT:672127","NEWT:656061","NEWT:37965","NEWT:656064","NEWT:3760","NEWT:36630","NEWT:381198","NEWT:1176302","NEWT:418106","NEWT:10029","NEWT:913645","NEWT:641809","NEWT:44250","NEWT:77151","NEWT:317447","NEWT:698761","NEWT:5530","NEWT:868565","NEWT:135674","NEWT:1562698","NEWT:11308","NEWT:12637","NEWT:2164133","NEWT:1180","NEWT:2767846","NEWT:1343074","NEWT:314344","NEWT:108061","NEWT:143292","NEWT:2767842","NEWT:314345","NEWT:224308","NEWT:3134131","NEWT:49928","NEWT:160621","NEWT:110368","NCBITaxon:5204","NEWT:1916073","NEWT:1303873","NEWT:1310161","NEWT:1230383","NEWT:77133","NEWT:160620","NCBITaxon:77643","NEWT:1310165","NEWT:114727","NEWT:5516","NEWT:498211","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:1410384","NEWT:1310173","NEWT:1072569","NEWT:1072568","NEWT:161934","NEWT:1148","NEWT:5508","NEWT:135666","NEWT:419481","NEWT:5507","NEWT:1140","NCBITaxon:2157","NEWT:2066563","NEWT:1143","NEWT:1336794","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:709964","NEWT:3710","NEWT:3712","NEWT:3711","NEWT:3714","NEWT:65121","NEWT:3716","NEWT:3715","NEWT:2065263","NEWT:1549675","NEWT:177437","NEWT:9014","NEWT:10891","NEWT:118221","NEWT:32201","NEWT:34869","NEWT:226126","NEWT:1616117","NEWT:34865","NEWT:28174","NEWT:550537","NEWT:284812","NEWT:284811","NEWT:3701","NEWT:3702","NEWT:1245466","NEWT:3704","NEWT:1246791","NEWT:34875","NEWT:10882","NEWT:1431297","NEWT:9005","NEWT:34871","NEWT:9006","NEWT:33548","NEWT:1883426","NEWT:657321","NEWT:1423687","NEWT:657322","NEWT:10886","NEWT:3707","NEWT:3708","NEWT:536231","NEWT:219813","NEWT:1301262","NEWT:319224","NEWT:2305479","NEWT:215451","NEWT:460519","NEWT:272843","NEWT:272844","NEWT:1348799","NEWT:55784","NEWT:485","NEWT:223074","NEWT:2499525","NEWT:467144","NEWT:142786","NEWT:28196","NEWT:2853422","NEWT:487","NEWT:1679718","NEWT:387344","NEWT:657318","NEWT:480","NEWT:67767","NEWT:46835","NEWT:216778","NEWT:376686","NEWT:237610","EFO:0001352","NEWT:219809","NEWT:219808","NEWT:119562","NEWT:1883446","NEWT:1233435","NEWT:118223","NEWT:141469","NEWT:272831","NEWT:987059","NEWT:491","NEWT:422203","NEWT:460523","NEWT:2422","NEWT:2423","NEWT:3755","NEWT:85694","NEWT:85692","NEWT:911045","NEWT:3750","NEWT:1195466","NEWT:2483192","NEWT:85698","NEWT:291146","NEWT:233271","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:278161","NEWT:415978","NEWT:410289","NEWT:191701","NEWT:9054","NEWT:472","NEWT:29459","NEWT:34839","NEWT:476","NEWT:28128","NEWT:470","NEWT:41122","NEWT:1328300","NEWT:748693","NEWT:106299","NCBITaxon:50557","NEWT:63363","NEWT:3735","NEWT:29491","NEWT:911022","NEWT:446","NEWT:153481","NEWT:714485","NEWT:445","NEWT:427924","NEWT:2003758","NEWT:29486","NEWT:63366","NEWT:63367","NEWT:29488","NEWT:215402","NEWT:3726","NEWT:1120755","NEWT:1862567","NEWT:591090","NEWT:30455","NEWT:28141","NEWT:9030","NEWT:9031","NEWT:3728","NEWT:29471","NEWT:108931","NEWT:219799","NEWT:1325292","NEWT:219798","NEWT:548","NEWT:150475","NEWT:546","NEWT:1431340","NEWT:256737","NEWT:51350","NEWT:8694","NEWT:8692","NEWT:542","NEWT:394503","NEWT:13076","NEWT:219785","NEWT:451515","NEWT:8673","NEWT:550","NEWT:10804","NEWT:52694","NEWT:554","NEWT:451516","NEWT:552","NEWT:1325291","NEWT:42410","NEWT:40686","NEWT:1225786","NEWT:366497","NEWT:771870","NEWT:8665","NEWT:51338","NEWT:771875","NEWT:377142","NEWT:9995","NEWT:28112","NEWT:49012","NEWT:28110","NEWT:10832","NEWT:311339","NEWT:507601","NEWT:520","NEWT:64657","NEWT:186441","NEWT:2585209","NEWT:643680","NEWT:214092","NEWT:40697","NEWT:13095","NEWT:162425","NEWT:242124","NEWT:9986","NEWT:8654","NEWT:536","NEWT:8658","NEWT:1268063","NEWT:8657","NEWT:9983","NEWT:8655","NEWT:80833","NEWT:479096","NEWT:28108","NEWT:28104","NEWT:1114957","NEWT:568703","NEWT:568708","NEWT:480020","NEWT:51315","NEWT:52641","NEWT:353152","NEWT:116597","NEWT:353153","NEWT:116598","NEWT:227377","NEWT:1774284","NEWT:27689","NEWT:207769","NEWT:40674","NEWT:51329","NCBITaxon:1763","NEWT:443906","NEWT:6055","NEWT:13037","NEWT:518","NEWT:519","NEWT:941638","NEWT:511","NEWT:7394","NEWT:6063","NEWT:404330","NEWT:1328388","NEWT:13064","NEWT:529120","NEWT:7375","NEWT:300641","NEWT:39655","NEWT:38323","NEWT:206411","NEWT:80869","NEWT:214053","NEWT:80863","NEWT:6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