Baricitinib and Lonafarnib Synergistically Target Progerin and Inflammation, Improving Lifespan and Health in Progeria Mice

Abstract

Hutchinson-Gilford progeria syndrome (HGPS) is a rare, fatal, and premature aging disorder caused by progerin, a truncated form of lamin A that disrupts nuclear architecture, induces systemic inflammation, and accelerates senescence. While the farnesyltransferase inhibitor lonafarnib extends the lifespan by limiting progerin farnesylation, it does not address the chronic inflammation or the senescence-associated secretory phenotype (SASP), which worsens disease progression. In this study, we investigated the combined effects of baricitinib (BAR), a JAK1/2 inhibitor, and lonafarnib (FTI) in a Lmna^G609G/G609G mouse model of HGPS. BAR + FTI therapy synergistically extended the lifespan by 25%, surpassing the effects of either monotherapy. Treated mice showed improved health, as evidenced by reduced kyphosis, better fur quality, decreased incidence of cataracts, and less severe dysgnathia. Histological analyses indicated reduced fibrosis in the dermal, hepatic, and muscular tissues, restored cellularity and thickness in the aortic media, and improved muscle fiber integrity. Mechanistically, BAR decreased the SASP and inflammatory markers (e.g., IL-6 and PAI-1), complementing the progerin-targeting effects of FTI. This preclinical study demonstrates the synergistic potential of BAR + FTI therapy in addressing HGPS systemic and tissue-specific pathologies, offering a promising strategy for enhancing both lifespan and health.

Keywords: Hutchinson–Gilford progeria syndrome; JAK-STAT; baricitinib; inflammation; lamin A; lifespan; progerin.

Figure 1

BAR + FTI therapy extends survival, enhances systemic health, and partially restores organ morphology in Lmna^G609G/G609G mice. (A): Kaplan–Meier graph showing the survival of Lmna^G609G/G609G mice across different treatment cohorts over time (MOCK: n = 28; BAR: n = 14; FTI: n = 13; BAR + FTI: n = 12). The MOCK HOM cohort showed the shortest lifespan, whereas the BAR + FTI combination displayed the most significant improvement in survival, surpassing both single treatments (BAR or FTI). (B): Radar chart illustrating systemic health parameters across different treatment cohorts. Each axis represents the degree of the effect observed for each health parameter. This chart allows for a direct visual comparison of health parameters. Larger enclosed areas indicate better systemic health. MOCK HOM mice showed the most severe deterioration in health parameters. The BAR + FTI-treated mice exhibited the most substantial overall improvements (MOCK: n = 28; BAR: n = 12; FTI: n = 13; BAR + FTI: n = 12). (C): Body weight changes over time in male (top plot) and female (bottom plot) mice. Color code is as shown in (A). All progeroid groups showed a progressive decline in body weight starting at 8–10 weeks of age (male: MOCK WT: n = 13; MOCK HOM: n = 11; BAR: n = 7; FTI: n = 6; BAR + FTI: n = 8; female: MOCK WT: n = 12; MOCK HOM: n = 18; BAR: n = 7; FTI: n = 7; BAR + FTI: n = 6). (D): Glucose tolerance was assessed using intraperitoneal glucose tolerance tests (ipGTTs) in male (top) and female (bottom) mice. MOCK HOM mice exhibited impaired glucose clearance. Glucose intolerance was further exacerbated in the BAR and BAR + FTI groups compared to that in the FTI alone group (males: MOCK WT: n = 8; MOCK HOM: n = 7; BAR: n = 5; FTI: n = 9; BAR + FTI: n = 8; females: MOCK WT: n = 9; MOCK HOM: n = 9; BAR: n = 6; FTI: n = 6; BAR + FTI: n = 7). (E): Quantification of glucose tolerance (ipGTT) as area under the curve (AUC) for male (top) and female (bottom) cohorts. Glucose tolerance was further reduced in the BAR and BAR + FTI cohorts but improved in the FTI single treatment (male: MOCK WT: n = 8; MOCK HOM: n = 7; BAR: n = 5; FTI: n = 9; BAR + FTI: n = 8; female: MOCK WT: n = 9; MOCK HOM: n = 9; BAR: n = 6; FTI: n = 6; BAR + FTI: n = 7). (F): Fasting blood glucose levels in male (top) and female (bottom) mice after a 6 h fasting period. All treatment groups with the Lmna^G609G/G609G genotype showed reduced fasting glucose levels compared with Lmna^+/+ mice (male: MOCK WT: n = 8; MOCK HOM: n = 7; BAR: n = 5; FTI: n = 9; BAR + FTI: n = 8; female: MOCK WT: n = 9; MOCK HOM: n = 9; BAR: n = 6; FTI: n = 6; BAR + FTI: n = 7). (G): Representative photographs at 90 days of age showing systemic health differences between the progeroid-treated groups. MOCK HOM mice showed severe kyphosis, fur abnormalities, and reduced body size. BAR and FTI treatments partially alleviated these symptoms, whereas the BAR + FTI treatment resulted in the most pronounced improvements. (H): Representative images of dissected organs at 90 days of age across the different cohorts. MOCK HOM mice exhibited a marked reduction in organ size, particularly in the spleen and thymus. FTI treatment further reduced the size of these organs, whereas BAR + FTI treatment improved their size and morphology. Detailed organ weight data are shown in Supplemental Figure S1. (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; ordinary one-way ANOVA test with Tukey’s post hoc test).

Figure 2

Combination therapy reduces STAT1/STAT3 activation and progerin accumulation in Lmna^G609G/G609G mice. (A,C,E): Representative images of western blots for P-STAT1, STAT1, P-STAT3, STAT3, lamin A/C, and progerin extracted from the aorta, skin, liver, and lung tissues of 90-day-old Lmna^+/+ and Lmna^G609G/G609G mice (B,D,F): Quantification of P-STAT1/STAT1 ratios (B), P-STAT3/STAT3 ratios (D), and progerin levels (F). Graphs show mean ± SD. (n = 3); * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ordinary one-way ANOVA test with Tukey’s post hoc test.

Figure 3

BAR + FTI combination improves progerin-induced tissue degeneration and fibrosis. (A,E,I,L): Histological analysis of skin (A), aorta (E), liver (I), and muscle (L) using hematoxylin and eosin (H,E, upper row) and Masson’s trichome staining (lower row), scale bar = 100 μm. In panel (A), HF denotes hair follicle, E epidermis, D dermis, S subdermis, M muscular layer. In panel (E), I denotes intima, M media, A adventitia. In panel (I), BV = blood vessel, I = interstitium and in (L) CN = central nucleus, S = sarcomere. MOCK-treated Lmna^G609G/G609G mice exhibited severe tissue degeneration and fibrosis across all tissues. (B–D): Quantification of skin parameters. Skin thickness in male (B) and female (C) mice is significantly reduced in MOCK HOM group, which was partially restored in BAR and BAR + FTI cohorts (male: MOCK WT: n = 3; MOCK HOM: n = 3; BAR: n = 3; FTI: n = 5; BAR + FTI: n = 4; female: MOCK WT: n = 5; MOCK HOM: n = 4; BAR: n = 3; FTI: n = 3; BAR + FTI: n = 5). (D): Dermal fibrosis is elevated in MOCK HOM mice and reduced in BAR + FTI cohorts compared to those receiving single treatments (MOCK WT: n = 7; MOCK HOM: n = 7; BAR: n = 9; FTI: n = 5; BAR + FTI: n = 8). (F–H): Quantification of aorta parameters. Media cellularity that is reduced in MOCK HOM mice is restored in BAR + FTI-treated animals (MOCK WT: n = 7; MOCK HOM: n = 7; BAR: n = 6; FTI: n = 6; BAR + FTI: n = 8). (G): Media thickness reduced in MOCK HOM animals is partially restored in BAR + FTI-treated mice (MOCK WT: n = 7; MOCK HOM: n = 8; BAR: n = 6; FTI: n = 6; BAR + FTI: n = 7). (H): Aortic media fibrosis, which is elevated in MOCK HOM- and FTI-treated mice, is reduced in BAR and FTI + FTI cohorts, with no marked improvement in FTI cohorts (MOCK WT: n = 6; MOCK HOM: n = 8; BAR: n = 6; FTI: n = 6; BAR + FTI: n = 6). (J,K): Hepatic tissue analysis. Increased cellularity in the MOCK HOM cohort, which is indicative of reduced hepatocyte size, is partially reduced in all treatment groups (MOCK WT: n = 3; MOCK HOM: n = 6; BAR: n = 6; FTI: n = 6; BAR + FTI: n = 6). (K): Fibrosis in hepatic tissue, which is most pronounced in the MOCK HOM- and FTI-treated groups, is significantly reduced in the BAR and BAR + FTI cohorts, with BAR + FTI showing the most efficacy (MOCK WT: n = 5; MOCK HOM: n = 6; BAR: n = 6; FTI: n = 6; BAR + FTI: n = 6). (M–O): Muscle tissue changes: Central nucleation is elevated in MOCK HOM mice and reduced in all treatment groups. Muscle fibrosis is pronounced in HOM MOCK and FTI mice but reduced in BAR + FTI mice. The sarcomere diameter is reduced in MOCK HOM mice and fully restored with FTI and BAR + FTI treatments (MOCK WT: n = 6; MOCK HOM: n = 6; BAR: n = 6; FTI: n = 6; BAR + FTI: n = 6) (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; ordinary one-way ANOVA test with Tukey’s post hoc test).

Figure 4

Heatmaps displaying differential mRNA expression across five treatment groups. Each profile represents the relative expression levels of the individual mRNA. Data are presented on a color scale ranging from 0 (dark purple = low expression) to 5 (yellow = high expression). The key markers analyzed include senescence-associated secretory phenotype (SASP), cytokines, and angiogenic and extracellular matrix (ECM)-related markers. The four organs examined are the (A) skin, (B) heart, (C) liver, and (D) spleen. The mRNA levels analyzed include IL-6, IL-8, IL-1a, IL-1b, IL-10, TNFα, PAI-1, TGF-1b, CXCL1, CCL2, IGFBP7, VCAM-1, CTGF, VEGF, and HIF-1. Fold changes are shown relative to the MOCK WT cohort. Data represent a minimum of n = 3 biological replicates per organ and treatment group.

Figure 5

Senescence, ECM remodeling, and inflammation in vascular and renal tissue. (A): Immunofluorescent staining in aorta sections at 90 days shows reduced αSMA in MOCK HOM tissue vs. MOCK WT; BAR and FTI monotherapies partially restored αSMA expression, while the BAR + FTI treatment nearly normalized it (n = 3). Lamin A/C (green), αSMA (red), and DNA (DAPI, blue) were detected. I denotes intima, M media and A adventitia. (B): Staining for the senescence marker p16 (green) and vimentin (red) in aorta sections. There was a high prevalence of p16-positive cells in the intima of HOM MOCK samples (indicated by arrows), which was persistent with FTI, reduced with BAR, and was nearly absent with BAR + FTI. Vimentin was decreased in MOCK HOM and partially restored by treatments (n = 3). (C): Aortic PAI-1 (green) and IL-6 (red) staining were elevated in MOCK HOM mice, especially in intima and adventitia (indicated by arrows). FTI had little effect, while BAR and BAR + FTI reduced both markers (n = 3). (D): Immunofluorescent staining in kidney glomeruli (G) showed high expression of senescence marker p16 (green) and reduced vimentin (red) in MOCK HOM mice. BAR and BAR + FTI reduced p16 and increased vimentin; FTI only increased vimentin (n = 3). Scale bars: (A–C): 100 µm; (D): 20 µm; blue color represents DAPI counterstaining of nuclei.