{"doi":"10.1101/2025.05.25.655991","title":"Multiplexed Targeted Spatial Mass Spectrometry Imaging Assays to monitor lipids and NAD <sup>+</sup> metabolites in male CD38 knockout mice exhibiting improved metabolism","abstract":"Abstract Mass spectrometry imaging (MSI) is a rapidly advancing technology that provides mapping of the spatial molecular landscape of tissues for a variety of analytes. Matrix-assisted laser desorption/ionization (MALDI)-MSI is commonly employed, however, confident in situ identification and accurate quantification of analytes remain challenging. We present a novel imaging methodology combining trapped ion mobility spectrometry (TIMS)-based parallel accumulation-serial fragmentation (PASEF) with MALDI ionization for targeted imaging parallel reaction monitoring (iprm-PASEF). We investigated the spatial distribution of lipids and metabolites in liver tissues from male wild-type and CD38 knockout mice (CD38 -/- ). CD38, an enzyme involved in nicotinamide adenine dinucleotide (NAD⁺) metabolism, significantly influences liver metabolic function and contributes to age-related NAD⁺ decline. Although CD38 deletion previously was linked to improved metabolic phenotypes, the underlying spatial metabolic mechanisms are poorly understood. The spatial iprm-PASEF workflow enabled confident identification and differentiation of lipid isomers at the MS2 fragment ion level and confirmed increased NAD + and decreased adenosine diphosphate ribose (ADPR), a by-product of NAD + hydrolysis, in CD38 -/- livers. This approach provided confident, specific, and robust MS2-based identification and quantification of fragment ions in spatial MSI experiments. Additionally, the innovative iprm-PASEF opens unprecedented opportunities for spatial metabolomics and lipidomics, offering spatially resolved insights into molecular mechanisms.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2025,"id":555001,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":5,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9523,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":814043,"name":"Joanna Bons","orcid":"0000-0002-1110-4193","position":1,"is_corresponding":false},{"id":1232987,"name":"Prasanna Vadhana Ashok Kumaar","orcid":null,"position":2,"is_corresponding":false},{"id":1232647,"name":"Jingqi Fang","orcid":"0000-0002-5733-4373","position":3,"is_corresponding":false},{"id":880957,"name":"Andrea Roberts","orcid":"0000-0003-0104-9124","position":4,"is_corresponding":false},{"id":1453286,"name":"Genesis Vega Hormazabal","orcid":null,"position":5,"is_corresponding":false},{"id":228953,"name":"Rebeccah Riley","orcid":"0000-0002-5870-6837","position":6,"is_corresponding":false},{"id":672984,"name":"Nannan Tao","orcid":null,"position":7,"is_corresponding":false},{"id":95315,"name":"Eric Verdin","orcid":"0000-0003-3703-3183","position":8,"is_corresponding":false},{"id":106307,"name":"Birgit Schilling","orcid":"0000-0001-9907-2749","position":9,"is_corresponding":false},{"id":341897,"name":"Charles A. Schurman","orcid":"0000-0002-5739-8481","position":0,"is_corresponding":true}],"reference_count":39,"raw_metadata":null,"created_at":"2026-07-19T02:54:54.542303Z","pmid":"40501674","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}