Aging poses a major risk factor for cardiovascular diseases, the leading cause of death in the aged population. However, the cell type-specific changes underlying cardiac aging are far from being clear. Here, we performed single-nucleus RNA-sequencing analysis of left ventricles from young and aged cynomolgus monkeys to define cell composition changes and transcriptomic alterations across different cell types associated with age. We found that aged cardiomyocytes underwent a dramatic loss in cell numbers and profound fluctuations in transcriptional profiles. Via transcription regulatory network analysis, we identified FOXP1, a core transcription factor in organ development, as a key downregulated factor in aged cardiomyocytes, concomitant with the dysregulation of FOXP1 target genes associated with heart function and cardiac diseases. Consistently, the deficiency of FOXP1 led to hypertrophic and senescent phenotypes in human embryonic stem cell-derived cardiomyocytes. Altogether, our findings depict the cellular and molecular landscape of ventricular aging at the single-cell resolution, and identify drivers for primate cardiac aging and potential targets for intervention against cardiac aging and associated diseases.
Aged ; Animals ; Humans ; Aging/genetics* ; Forkhead Transcription Factors/metabolism* ; Myocytes, Cardiac/metabolism* ; Primates/metabolism* ; Repressor Proteins/metabolism* ; Transcriptome ; Macaca fascicularis/metabolism*

Cell Cycle Proteins ; Microtubule-Associated Proteins

Gallic Acid/pharmacology* ; Humans ; Senotherapeutics ; Stem Cells

Animals ; Chloroquine/pharmacology* ; Longevity ; Rats

OBJECTIVE：To provide reference f or the qualit y sta ndard establishment of Amaranthus retroflexus. METHODS ： Taking 7 batches of A. retroflexus medicinal materials as the research object ，the appearance properties of the medicinal materials were investigated ，and the microscopic characteristics of the medicinal powders were observed. TLC method was adopted to qualitatively identify rutin ，valine and leucine in A. retroflexus medicinal materials. According to the relevant methods of the 2015 edition of Chinese Pharmacopoeia （part Ⅳ），water content ，total ash content ，acid-insoluble ash content and water-soluble extract content were determined. HPLC method was used to determine the content of rutin in the medicinal material of A. retroflexus . The determination was performed on Agilent 5 TC-C18（2）column with mobile phase consisted of methanol- 0.3％ phosphoric acid solution（40∶60，V/V），at the flow rate of 1.0 mL/min. The detection wavelength was set at 358 nm，and the column temperature was 30 ℃. The sample size was 10 μL. RESULTS：The appearance and microstructure characteristics of the medicinal materials were consistent with the existing description. The identification results of TLC meth od showed that 7 batches of medicinal materials and each reference substance （rutin，valine，leucine）showed spots of the same color at the same position. The moisture content of 7 batches of A. retroflexus medicinal materials was 7.43％-8.72％，the total ash content was 11.82％-13.78％，the acid-insoluble ash content was 0.15％-0.55％，and the water-soluble extract content was 17.27％-24.74％. The linear range of rutin was 10-200 μg/mL（R 2＝1.000 0）. RSDs of precision test ，stability test （24 h）and repeatability test were all less than 2.0％ （n＝6）. The average recovery rates of rutin were 99.14％，97.98％ and 98.80％ in low ，medium and high concentration of samples，and RSDs were 0.97％，0.95％，0.96％（n＝3）. The contents of rutin in 7 batches of A. retrophylla were 0.314-1.102 mg/g. CONCLUSIONS：In this study ，character observation ，microscopic identification ，moisture content ，total ash content ，acid- insoluble ash content and water-soluble extract content of A. retroflexus are investigated ；TLC method was established for qualitative identification of leucine ，valine and rutin in A. retroflexus ，and the HPLC method was established for content determination of rutin. It provides reference for the quality standard establishment of A. retroflexus .

Aging/metabolism* ; Animals ; Gene Expression Regulation ; Macaca fascicularis ; Retina/metabolism* ; Single-Cell Analysis

Many human genetic diseases, including Hutchinson-Gilford progeria syndrome (HGPS), are caused by single point mutations. HGPS is a rare disorder that causes premature aging and is usually caused by a de novo point mutation in the LMNA gene. Base editors (BEs) composed of a cytidine deaminase fused to CRISPR/Cas9 nickase are highly efficient at inducing C to T base conversions in a programmable manner and can be used to generate animal disease models with single amino-acid substitutions. Here, we generated the first HGPS monkey model by delivering a BE mRNA and guide RNA (gRNA) targeting the LMNA gene via microinjection into monkey zygotes. Five out of six newborn monkeys carried the mutation specifically at the target site. HGPS monkeys expressed the toxic form of lamin A, progerin, and recapitulated the typical HGPS phenotypes including growth retardation, bone alterations, and vascular abnormalities. Thus, this monkey model genetically and clinically mimics HGPS in humans, demonstrating that the BE system can efficiently and accurately generate patient-specific disease models in non-human primates.
Animals ; Disease Models, Animal ; Female ; Gene Editing ; Humans ; Lamin Type A/metabolism* ; Macaca fascicularis ; Progeria/pathology*