The Long Life Family Study (LLFS) has enrolled 4,953 participants in 539 pedigrees in the USA and Denmark that are enriched for exceptional longevity, and has measured them longitudinally in three extensive in-person visits measuring key healthy aging phenotypes in all of the major domains of the aging process. We have demonstrated through many publications that selecting on longevity in the first (proband) generation, results in the second (offspring) generation being much healthier than average in many key phenotypes. Analysis of Danish Medical Registry data suggested that the protection persists into this third generation. In Visit 3, we expanded our LLFS pedigrees to include grandchildren. However, LLFS pedigrees are heterogeneous by phenotype, with different families showing familial clustering of protection in cognition, grip strength, pulmonary function, blood pressure, etc. Further linkage analysis identified extremely strong genetic linkage peaks for cross-sectional as well as longitudinal trajectory rates of change phenotypes for a wide variety of healthy aging domains such as exceptional cognitive performance and resilience to Alzheimer's Disease and Related Dementias (ADRD). Pedigree specific LODs and Whole Genome Sequencing (WGS) suggests that these peaks are driven by rare, protective variants running in selected pedigrees. Through linkage and genomc-wide analysis, we found many new rare and protective variants for healthy aging phenotypes (HAPs), including a new gene for AD, MTUS2. On a subset of the cohort (pedigrees with strong linkage evidence), we generated extensive longitudinal OMICs (transcriptomics, methylomics and metablomics). Despite our success, LLFS is almost all European Ancestry (EA). Adding African-Ancestry (AA) pedigrees will allow us to find new protective variants that are unlikely to be found in EAs (because of genetic drift), and to better resolve the causal variants common to both (because the haplotype blocks of AAs are only half as large as EAs reducing the “driver vs. passenger problem”). Visit 1 was nearly 20 years ago. Since then, there have been many medical advances and healthcare changes. We need to add a modest number of concurrent EA families, to contrast with the new AA ones, to avoid the confounding of secular trends with ancestry differences. The offspring are now approaching 80 years old on average, when expected disease incidence increases. Extending longitudinal follow-up of the original pedigrees will better define HAP trajectories, giving greater power to find protective variants for EL and HAPs. We will extend characterization of LLFS to additional domains microbiome, somatic mutation (CHIP), proteomics. To examine associated haplotypes and structural variants, long-read sequencing is crucial. 70% of human structural variation found in long-read WGS is missed by short-read WGS. Extending the omics to the full cohort will help us understand mechanisms of action of associated variants to protective EL and HAPs. Finally, a more sophisticated systems biology approach, including using state of the art Artificial Intelligence approaches, are needed to fully understand the role of genes working in networks to produce HAPs and EL.
