Science - USA (2022-02-11)

674 11 FEBRUARY 2022¥VOL 375 ISSUE 6581 science.orgSCIENCE

A

URAD ARHGDIG

ANKDD1A

KRT5

TNMDCES1

SPARCASAH1

CLU

P2RX6 CA3

FRG1−DT

UCHL1 PAIP2B

NDRG4

LOX

DHDDS

SEMA4B

0.0

2.5

5.0

7.5

10.0

−2 −1 0 1 2

log2FoldChange

−log10(padj)

12m vs Baseline

CLU

UCHL1C RYA B CA3

TNMD

URAD

RSPO1 AASS

ANKDD1A

IL1RAP

CALCRL

HSD11B1 PERP

CFH

P2RX6

ACVR1C

S100A4

PDE3A

0

3

6

9

−4 −2 0 2

log2FoldChange

−log10(padj)

24m vs Baseline

padj < 0.05 not significant

E

C

MHC class II antigen presentation (R)

Lysosome (K)

Complement cascade (R)

IFNg signaling (R)

DECTIN1 mediated non-canonical NFkB (R)

IL5 pathway (B)

Core matrisome (N)

Proton coupled monocarboxylate transport (R)

Integration of energy metabolism (R)

Sumoylation (R)

Insulin signaling pathway (K)

Transcriptional activation of mitochondrial biogenesis (R)

PPARA pathway (B)

BMAL1 clock NPAS2 activates circadian gene expression (R)

−2 −1 0 1 2

NES

Pathway analysis: 24m vs Baseline

F

24

11.50

11.75

12.00

12.25

12.50

Baseline 12 24

Expression level (FMO2)

padj = 0.006

0.0

0.1

0.2

0.3

Baseline 12

Fraction of macrophages

p = 0.04

p = 0.04

G

D

rank

0

2500

5000

7500

10000

12500

0.0 0.2 0.4 0.6

enrichment score

PPARA pathway

NES = 2.03

padj = 0.018

0

2500

5000

7500

10000

12500

0.0 0.2 0.4 0.6

enrichment score

Transcriptional activation

of mitochondria biogenesis

NES = 1.95

padj = 0.018

0

2500

5000

7500

10000

12500

−0.5 −0.4 −0.3 −0.2 −0.1 0.0

enrichment score

Lysosome

NES = -1.98

padj = 0.018

B GSE20536: twin-pairs discordant for physical

activity for over 30 years

0.0 -0.2 -0.4 -0.6 -0.8

enrichment score

0

10000

20000

30000

40000

50000

0.6 0.4 0.2 0.0

enrichment score

rank

Up in CALERIE

(24m vs baseline)

NES = 1.55

p = 0.002

Down in CALERIE

(24m vs baseline)

NES = -1.78

p = 0.0004

activenon-active

SPP1

CHIT1

MMP9

TREM2

MSR1

PLA2G7

CCL19

VSIG4

SIGLEC1

CHI3L2

CD163

CHI3L1

C1QC

APOC1

IL1RN

C1QB

LILRB5

FPR3

DHRS9

ST14

2 1 0 −1 −2

baseline 12 month 24 month

z-score

Fig. 2. CR in humans reveals anti-aging immunometabolic checkpoints.
(A) Volcano plots depicting the results of differential gene expression analysis of
adipose tissue. Each dot represents a gene. Top 18 significantly up- or down-
regulated genes at 1 year (left) or 2 years (right) of CR compared with baseline
(n= 8). (B) Gene set enrichment analysis (GSEA) of genes up- and down-regulated
after 2 years of CR in a signature describing differences between active and
nonactive twins (GSE20536). (C) Selected pathways significantly (FDR, 5%)
regulated by 2 years of CR based on GSEA. Red and blue bars indicate pathways
up- and down-regulated by CR, respectively. Letters correspond to the source of

pathway gene set. K, KEGG; R, Reactome; B, BioCarta. (D) Enrichment curves for

selected significantly regulated pathways.y-axis denotes rank of the gene in

list ordered by log2FC. (E) Changes in the expression level of FMO-2 after CR. Adjusted

Pvalues (padj) were calculated in the differential gene expression analysis. (F) Box

plots showing estimated macrophage fractions after deconvolution of adipose

transcriptome in adipose tissue at baseline and 1 and 2 years after CR.Pvalues

were calculated using the pairedttest. (G) Expression of genes specific to adipose

tissueÐresident macrophages shown at baseline and 1 and 2 years after CR. NES

corresponds to the normalized enrichment score.

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