A Non-targeted Proteomics Newborn Screening Platform for Genetic Disorders

Newborn screening using dried blood spot (DBS) samples has made a substantial contribution to public healthcare by detecting patients with genetic disorders as neonates. Targeted measurements of nucleic acids and metabolites have played major roles in newborn screening to date, while the feasibility of new non-targeted approaches, including genome-wide DNA sequencing, has been explored. Here, we investigated the applicability of non-targeted quantitative proteomics analysis to newborn screening for genetic diseases. DBS protein profiling allowed monitoring of levels of proteins encoded by 2912 genes, including 1106 listed in the Online Mendelian Inheritance in Man database, in healthy newborn samples, and was useful in identifying patients with inborn errors of immunity by detecting reduced levels of disease causative proteins and cell-phenotypical alterations. Our results indicate that application of non-targeted quantitative protein profiling of DBS samples can forge a new path in screening for genetic disorders. ### Competing Interest Statement The authors have declared no competing interest. ### Funding Statement JSPS KAKENHI under Grant Numbers 19K17328 and 21K07795; by AMED under Grant Number JP23ek0109586; and by the Research on Measures for Intractable Diseases project from the Japanese Ministry of Health, Labor, and Welfare. ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: Ethics committee of Kyoto University Hospital gave ethical approval for this work. I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes All of the data used to generate the figures are available in the published article and its online supplemental material. * CHS : Chédiak-Higashi syndrome DBS : dried blood spot DEPs : differentially expressed proteins DIA : data-independent acquisition FHL : familial hemophagocytic lymphohistiocytosis GO : Gene Ontology HPS : Hermansky-Pudlak syndrome IEIs : inborn errors of immunity IEMs : inborn errors of metabolism KRECs : kappa-deleting element recombination circles LC-MS/MS : liquid chromatography-assisted mass spectrometry NBS : newborn screening OMIM : Online Mendelian Inheritance in Man TRECs : T cell receptor excision circles WAS : Wiskott-Aldrich syndrome XLA : X-linked agammaglobulinemia