TCONS_00052110 is associated with prolonged PTBP1 half-life and mitochondrial injury in PAH-associated right ventricular stress

Xiaowei Gao; Yue Yang; Lizhe Guo; Lu Wang; Li Qian; Gang Qin; Hui Luo; Yanan Cao; E Wang

doi:10.1038/s41598-026-50545-8

. 2026 Apr 29;16:19892. doi: 10.1038/s41598-026-50545-8

TCONS_00052110 is associated with prolonged PTBP1 half-life and mitochondrial injury in PAH-associated right ventricular stress

Xiaowei Gao ¹, Yue Yang ¹, Lizhe Guo ¹, Lu Wang ¹, Li Qian ¹, Gang Qin ¹, Hui Luo ¹, Yanan Cao ^1,^✉, E Wang ^1,^2,^✉

PMCID: PMC13315928 PMID: 42056443

Abstract

Right ventricular failure (RVF) is the major cause of mortality in pulmonary arterial hypertension (PAH), and even mild inflammatory stress can precipitate rapid decompensation. Here we report that the long noncoding RNA TCONS_00052110 (TCONS) is upregulated in the right ventricle (RV) under inflammatory stress and may modulate stress-associated responses. In adult male Sprague–Dawley rats with monocrotaline-induced PAH, a low-dose lipopolysaccharide challenge precipitated acute RVF. Mechanistically, TCONS physically associates with polypyrimidine tract–binding protein 1 (PTBP1) and is associated with a prolonged PTBP1 protein half-life, consistent with reduced PTBP1 protein turnover. Elevated PTBP1 skews pyruvate kinase muscle (PKM) isoforms toward PKM2, favoring a PKM2-dominant metabolic state consistent with glycolysis-related remodeling. These changes are accompanied by mitochondrial injury and cytosolic cytochrome c release in vivo and in vitro. Using a cardiomyocyte-enriched AAV9-cTnT strategy, knockdown of TCONS was associated with normalization of the PKM2/PKM1 balance, attenuation of mitochondrial injury, preservation of RV functional indices after inflammatory challenge, and improved survival. Reanalysis of patient-derived datasets from PAH lung tissue and RV tissue spanning compensation-to-decompensation revealed enrichment of inflammatory and glycolysis-related pathways concordant with the TCONS–PTBP1 axis, supporting contextual relevance. These findings support a model of a post-transcriptional link between inflammatory stress, glycolysis-related remodeling, and mitochondrial injury in PAH-related RVF, warranting validation in human RV tissue.

Supplementary Information

The online version contains supplementary material available at 10.1038/s41598-026-50545-8.

Keywords: Pulmonary arterial hypertension (PAH), Right ventricular failure (RVF), Inflammatory stress (LPS), TCONS_00052110 (TCONS), PTBP1, Glycolysis-related remodeling, Mitochondrial injury

Subject terms: Cardiology, Cell biology, Diseases, Medical research, Physiology

Introduction

Patients with pulmonary arterial hypertension (PAH) exhibit a marked clinical vulnerability in which even minor systemic inflammatory stress—such as mild infections or pregnancy-related physiological stress, without overt sepsis—can precipitate abrupt right ventricular (RV) decompensation and life-threatening right ventricular failure (RVF)^1–4. Yet the molecular mechanisms by which inflammatory cues precipitate energetic collapse in the PAH RV remain incompletely defined.

Mitochondrial injury is increasingly recognized as a central pathological mechanism in RVF^2,5. Under stress conditions, impaired oxidative phosphorylation (OXPHOS) results in excessive accumulation of mitochondrial reactive oxygen species (mtROS), disrupts redox homeostasis, and activates inflammatory cascades such as NLRP3 inflammasome signaling, while disruption of the mitochondrial membrane leads to cytosolic release of cytochrome c and amplification of sterile inflammation and innate immune activation^6–9. These processes are closely associated with a PKM2-dominant metabolic state, including features consistent with reduced mitochondrial respiration and increased reliance on glycolysis, which is a hallmark of stressed cardiomyocytes and has been linked to maladaptive RV remodeling in decompensated PAH^10,11. The resultant changes encompass glycolysis-related remodeling, impaired OXPHOS, and redox imbalance.

The RNA-binding protein polypyrimidine tract-binding protein 1 (PTBP1) plays a key role in regulating alternative splicing of pyruvate kinase muscle (PKM) isoforms, promoting PKM2 expression and shifting cellular metabolism toward glycolysis^12,13. Increased dimeric PKM2 in cardiomyocytes has been associated with reduced mitochondrial respiration and energetic failure¹⁴. Moreover, upstream regulators that couple inflammatory stress to PTBP1–PKM2 signaling in the PAH RV have not been defined.

To determine whether PTBP1–PKM2 signaling contributes to mitochondrial dysfunction and acute RVF, we established a two-hit model by combining monocrotaline (MCT)-induced PAH with lipopolysaccharide (LPS)-induced systemic inflammation to mimic clinical inflammatory triggers^15,16. By examining molecular alterations in the RV myocardium, we identified long noncoding RNAs (lncRNAs) as potential regulators of inflammatory and metabolic pathways¹⁷. High-throughput sequencing revealed that TCONS_00052110 (TCONS), also known as lncRNA XLOC_034025, is a novel lncRNA upregulated in the RV myocardium of PAH rats following LPS injection¹⁸. Emerging evidence suggests that lncRNAs can modulate cardiac remodeling and heart failure progression^19,20. However, it remains unclear whether TCONS regulates the PTBP1–PKM2 axis or contributes to mitochondrial and energy metabolism in cardiomyocytes.

In this study, we used an LPS two-hit PAH model to investigate the pathophysiology of acute RVF under inflammatory stress. Through in vivo and in vitro approaches, we explored the role of TCONS in regulating myocardial mitochondrial function, specifically its association with PTBP1, its effect on the PKM2/PKM1 ratio, and the downstream consequences for RV mitochondrial integrity and mitochondrial injury-related phenotypes under inflammatory challenge.

Results

Inflammatory stress precipitates acute right ventricular failure in PAH accompanied by glycolysis-related remodeling and mitochondrial injury

Our previous research showed that the PAH right ventricle is highly sensitive to inflammation, with even mild stimuli rapidly triggering an inflammatory cascade and RVF¹⁵. Building on these findings, we further investigated the metabolic and molecular mechanisms underlying this vulnerability and examined their relevance in human disease.To this end, we confirmed in rats that low-dose systemic inflammatory stimulation rapidly induced RVF in PAH rats, whereas healthy controls were unaffected. Consistent with our previous validation of this model^16,21, echocardiographic parameters did not differ significantly between Control and Control + LPS groups, indicating that this LPS dose does not independently impair right ventricular function in otherwise healthy rats. Echocardiographic analysis revealed significant reductions in TAPSE and increases in RVEDD in PAH + LPS rats, accompanied by markedly decreased strain rates of both the RV free wall and interventricular septum (Fig. 1A–D). HR, LVCO, and LVSV were all decreased, with increased sPAP but no change in mid-RV strain rate (Fig. S1), supporting the presence of acute RVF specifically in inflamed PAH rats.

TEM showed severe mitochondrial swelling, cristae disruption, and reduced clustering in the right ventricle of RVF rats (Fig. 1E,F). These ultrastructural alterations were accompanied by marked inhibition of mitochondrial oxidative phosphorylation (OXPHOS) and increased cytochrome c release (Figs. 1G, 2A,B), indicating that even mild inflammation can precipitate profound metabolic disturbance in RVF.

To determine whether these findings are relevant to human disease, we reanalyzed patient-derived transcriptomic datasets from PAH lung tissue and human right ventricular tissue spanning compensated-to-decompensated states (Figs. S2, S3). Transcriptomic analysis of PAH lung tissues (Fig. S2A) revealed robust separation between PAH and control samples by principal component analysis (PCA). The top 50 differentially expressed genes (DEGs) effectively distinguished samples by disease status, and gene set enrichment analysis (GSEA) confirmed significant activation of immune response, phagocytosis, and lysosomal pathways in PAH lungs, supporting an important role for metabolic-inflammatory dysregulation in PAH pathogenesis (Fig. S2B–D). In RV tissue datasets spanning control, compensated, and decompensated (RVF) patients (Fig. S3), transcriptomic profiling revealed progressive alterations in metabolic and inflammatory pathways with RVF progression. Volcano plots and unsupervised DEG clustering showed clear molecular distinctions between groups, with pathway analysis highlighting upregulation of glycolysis, hypoxia responses, mitochondrial function-related programs, and immune activation in decompensated RVF. Ridge and dot plots further illustrated progressive dysregulation of mitochondrial and inflammatory pathways as RV function worsened (Fig. S3C–F).

We further confirmed in RVF rats that PTBP1 protein expression was upregulated without corresponding changes in mRNA (Fig. 2C), suggesting post-transcriptional regulation. Simultaneously, both PKM1 protein and mRNA levels were reduced, while PKM2 protein and mRNA levels increased, resulting in a significantly elevated PKM2/PKM1 ratio (Fig. 2D, Fig. S1A–C), indicating a PKM2-dominant state consistent with glycolysis-related remodeling under inflammatory stimulation.

Collectively, by integrating patient-derived transcriptomic and experimental data, we show that preexisting PAH renders the right ventricle highly susceptible to even mild inflammatory insults, resulting in acute functional decompensation. This process is associated with molecular and respiratory features consistent with reduced OXPHOS and glycolysis-related remodeling, supporting a model in which immune–metabolic coupling is a biologically relevant associated component of acute RVF.

LncRNA TCONS_00052110 is induced in RVF rats and is characterized as a novel full-length noncoding transcript with post-transcriptional regulatory potential

We observed that the TCONS (TCONS_00052110) qRT-PCR signal was significantly increased by inflammatory stimulation, especially in the PAH group (Fig. 3A), supporting induction of this transcript at the disease stage associated with acute RV decompensation. RNA-FISH detected a predominantly cytoplasmic signal consistent with TCONS localization of cardiomyocytes under inflammatory conditions (Fig. 3B), indicating a role in post-transcriptional regulation.

Fig. 3 — Characterization and predicted PTBP1 interaction of TCONS in RVF. (A) RV TCONS expression by qRT–PCR in Control, Control + LPS, PAH, and PAH + LPS. Each symbol represents one rat (n = 3/group); data are mean ± SD. (B) RNA-FISH showing cytoplasmic localization of TCONS in cardiomyocytes (DAPI counterstain). Scale bar: 20 µm. (C) Predicted secondary structure of full-length TCONS (2536 nt). (D, E) Coding-potential analyses (CPC/CNCI) and ORF distribution. (F–H) Predicted PTBP1-binding motifs, RPISeq interaction probability, and conservation with conserved sequence blocks aligned to predicted binding regions. Panels B–H show representative outputs from ≥ 3 independent analyses. Statistics: (A): ordinary two-way ANOVA (PAH × LPS): interaction P = 0.0009; PAH P = 0.0006; LPS P < 0.0001 (Tukey post hoc as indicated). ns, not significant; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

To define the transcript more directly, we performed RACE using RV myocardial tissue from PAH + LPS rats, i.e., the same disease context in which the TCONS signal was induced. This analysis identified a 2536-nt full-length transcript, as supported by Sanger sequencing and coverage analysis (Fig. S4A and Table S6). RNAfold-based secondary structure prediction revealed multiple stem–loop domains (Fig. 3C), consistent with the possibility that TCONS may function as a scaffold or interaction platform for RNA-binding proteins. Coding-potential analyses using CPC and CNCI supported interpretation of TCONS as a noncoding transcript (Fig. 3D). ORF analysis identified only short ORFs, none of which supported a conventional protein-coding transcript model.

Genomic mapping localized TCONS to rat chromosome 10:103,693,728–103,695,182 (Rnor 6.0), adjacent to the Socs3 gene cluster. No annotated lncRNA corresponding to this transcript was identified in the current Ensembl, GENCODE, or RefSeq rat databases, supporting the interpretation that TCONS represents a previously uncharacterized transcript at this locus (Fig. S4B). Homology searches in human and mouse identified syntenic regions without annotated lncRNA counterparts, supporting locus conservation and transcript novelty (Fig. S4C,D). Because this locus lies adjacent to and partially overlaps the Socs3 region, we interpret TCONS as a distinct noncoding transcript species at a complex locus, rather than as a conventional SOCS3 protein-coding isoform.

Motif and interaction analyses further suggested potential functional relevance of this transcript. Multiple PTBP1-binding motifs were predicted along the TCONS sequence (Fig. 3F), and RPISeq analysis identified two regions with high predicted interaction probability with PTBP1 (Fig. 3G). Several predicted binding regions overlapped with conserved sequence blocks (CSBs) (Fig. 3H), supporting the possibility that functionally relevant RNA–protein interaction features may be preserved across species. Together, these findings support the identification of TCONS_00052110 as a novel full-length lncRNA induced in the PAH + LPS RV, with structural and localization features consistent with a post-transcriptional regulatory role.

Inflammatory two-hit stress drives oxidative and mitochondrial injury in hypertrophic cardiomyocytes

To model inflammation-induced RVF, we established a two-hit cellular model by sequentially inducing hypertrophy and then applying inflammatory stimulation. H9C2 cardiomyocytes were treated with arginine vasopressin (AVP) for 72 h, resulting in a hypertrophic phenotype characterized by a dose-dependent increase in cytoplasmic α-actinin expression (Fig. 4A). Quantitative analysis showed that AVP at 0.1 and 1 µM elevated MYHCB expression by 1.37-fold and 1.58-fold, respectively, and TRPC6 expression by 5.3-fold and 7.8-fold, respectively, compared to controls (Fig. 4B). Based on previous research by Klein et al.²² and our own validation, 1 µM was selected as the optimal concentration for inducing hypertrophy, as further confirmed by western blotting (Fig. 4C).

Fig. 4 — A two-hit cellular model reveals enhanced susceptibility of hypertrophic cardiomyocytes to inflammatory stress. (A) α-actinin immunofluorescence after AVP (0.1 or 1 μM, 72 h) and quantification per cell. Scale bar: 20 µm. (B) qRT–PCR of hypertrophy markers TRPC6 and MYHCB after AVP. (C) α-actinin immunoblot and densitometry (Control vs AVP 1 μM). (D, E) ROS time course in normal vs hypertrophic H9C2 cells after LPS (DCFH-DA flow cytometry): quantification (D) and representative plots (E). (F) Representative MitoT-Rea (red), Cyto C (green), DAPI (blue). Scale bar: 20 µm. Zoom in representative merge images. Scale bar: 7.5 µm. (G) Intracellular Ca²⁺ (Fluo-3/AM) quantification; representative images in (I). Scale bar: 20 µm. (H) Cytochrome c signal quantification. (I) Representative Fluo-3/AM images. Each symbol represents one independent cell-culture experiment; technical replicates averaged within each experiment (n as indicated in panels). Data are mean ± SD. Statistics: A–B, one-way ANOVA with Tukey (overall P ≤ 0.0014; pairwise P values as indicated). C, unpaired two-tailed t-test P = 0.0218. D, two-way RM ANOVA: interaction P = 0.0068; time P = 0.0003; group P = 0.0091 (Šídák comparisons as indicated). G–H, two-way ANOVA: Ca²⁺ (G) interaction P < 0.0001; cytochrome c (H) interaction P = 0.0041 (post hoc as indicated). ns, not significant; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

We next evaluated oxidative stress under inflammatory stimulation. Flow cytometric analysis using the ROS-sensitive probe DCFH-DA demonstrated that LPS (100 μg/ml for 6 h) significantly increased ROS production in hypertrophic cardiomyocytes, but not in non-hypertrophic cells, as indicated by elevated mean fluorescence intensity (MFI) and a rightward shift in the FITC-A histogram (Fig. 4D,E). These results suggest that hypertrophic cells are more metabolically vulnerable to inflammatory stress.

To examine mitochondrial dynamics, we performed MitoTracker Red and cytochrome c (Cyto C) co-immunofluorescence staining. In the absence of LPS, both normal and hypertrophic cardiomyocytes displayed elongated mitochondrial morphology and cytoplasmic Cyto C retention. However, LPS challenge led to a dramatic transformation in hypertrophic cells, characterized by fragmented spherical mitochondria and increased cytosolic Cyto C signal, indicating mitochondrial destabilization and consistent with early apoptotic activation (Fig. 4F). Quantitative analysis of Cyto C fluorescence confirmed a significant increase in cytoplasmic Cyto C intensity in hypertrophic cardiomyocytes following LPS treatment, but not in their non-hypertrophic counterparts (Fig. 4G), highlighting selective mitochondrial vulnerability in the hypertrophic context.

Calcium imaging using Fluo-3/AM further demonstrated that LPS-induced calcium influx was markedly elevated in hypertrophic cells, whereas non-hypertrophic cells remained unaffected (Fig. 4H,I). This suggests that hypertrophic cardiomyocytes are predisposed to inflammatory calcium overload, potentially contributing to mitochondrial collapse.

Together, these data demonstrate that pre-existing cardiomyocyte hypertrophy renders cells highly susceptible to inflammatory injury via increased ROS production, mitochondrial dysfunction, calcium overload, and apoptotic priming. This two-hit model recapitulates key pathophysiological features of inflammation-driven RVF in the setting of PAH.

TCONS is associated with PTBP1 accumulation and reduced PTBP1 protein turnover in vitro

Given consistent upregulation of TCONS under inflammatory conditions in both in vivo and in vitro settings, we further investigated its functional role in cellular stress responses. RNA-FISH analysis revealed that TCONS predominantly localized to the cytoplasm of hypertrophic H9C2 cardiomyocytes treated with AVP and LPS (Fig. 5A), suggesting potential involvement in post-transcriptional regulation. Agarose gel electrophoresis confirmed successful synthesis of full-length TCONS transcripts in vitro (Fig. S5A).

Fig. 5 — TCONS physically associates with PTBP1 and is associated with reduced PTBP1 turnover under inflammatory stress. (A) RNA-FISH of TCONS in AVP + LPS-treated H9C2 cells (DAPI counterstain; 18S rRNA control as indicated). Scale bar: 5 µm. (B) Silver-stained SDS-PAGE of RNA pull-down eluates. (C–E) Mass spectrometry and bioinformatic analyses identifying PTBP1 as a high-confidence TCONS-interacting protein and enrichment of RNA-processing pathways. (F) Immunoblot validating PTBP1 enrichment in sense-strand pull-down vs antisense/bead controls. (G, H) Docking/3D modeling predicting PTBP1-binding motifs on TCONS. (I, J) Truncation mapping of PTBP1-binding regions. (K–L) CHX chase assays showing that TCONS knockdown accelerates, whereas TCONS overexpression prolongs, PTBP1 half-life. RNA pull-down and CHX chase were performed in three independent cell-culture preparations (n = 3 biological replicates) with consistent results; representative blots are shown.

To identify proteins associated with TCONS, we performed RNA pulldown assays using biotin-labeled sense and antisense TCONS transcripts. SDS-PAGE analysis showed several bands specifically enriched in the sense-strand group (Fig. 5B). Mass spectrometry combined with GO/KEGG enrichment analysis indicated that these candidate proteins were mainly involved in RNA splicing, RNA metabolism, and stress-response pathways. Among them, PTBP1 emerged as a prioritized candidate interactor (Fig. 5C–E, Fig. S5B–F, and Table S8). The association between TCONS and PTBP1 was further supported by sense-versus-antisense pulldown-based immunoblot validation (Fig. 5F).

To further explore the RNA–protein interaction interface, in silico docking analysis identified two candidate PTBP1-associated regions with high predicted interaction probability at nucleotides 2136–2237 and 2186–2287 of TCONS (Fig. 5G,H). We then synthesized four truncated TCONS fragments and evaluated PTBP1 enrichment by RNA pulldown. Deletion of either candidate region significantly reduced PTBP1 enrichment (Fig. 5I,J), supporting the relevance of these PTBP1-associated regions.

To determine whether TCONS affects PTBP1 protein turnover, we performed cycloheximide (CHX) chase assays. TCONS knockdown accelerated PTBP1 degradation, whereas TCONS overexpression prolonged PTBP1 protein half-life and delayed its loss (Fig. 5K,L). Together, these findings support a model in which TCONS physically associates with PTBP1 and is associated with reduced PTBP1 protein turnover, thereby favoring PTBP1 accumulation. Whether this effect reflects proteasomal or alternative degradation pathways requires further mechanistic validation.

TCONS knockdown attenuates mitochondrial injury and disrupts the PTBP1–PKM2 axis

To determine the functional consequence of TCONS silencing under inflammatory conditions, we confirmed effective knockdown of TCONS using two siRNAs in LPS-treated hypertrophic cardiomyocytes (Fig. 6A). PTBP1 mRNA levels remained unchanged (Fig. 6B), whereas PKM1 transcripts were upregulated and the PKM2/PKM1 mRNA ratio was significantly reduced following TCONS knockdown (Fig. 6C,D). Western blotting further demonstrated decreased PTBP1 and PKM2 protein expression, along with increased PKM1 levels, leading to a reduced PKM2/PKM1 ratio (Fig. 6E–H). These findings support a post-transcriptional mechanism through which TCONS is associated with PTBP1 accumulation and promotes a PKM2-dominant metabolic state consistent with glycolysis-related remodeling.

Fig. 6 — TCONS knockdown attenuates mitochondrial injury while disrupting the PTBP1–PKM2 axis under inflammatory stress. (A–D) qRT–PCR of TCONS (A), Ptbp1 (B), Pkm1 (C), and PKM2/PKM1 ratio (D) in AVP-induced hypertrophic H9C2 cardiomyocytes under AVP, AVP + LPS, AVP + NC + LPS, AVP + Si + LPS, and AVP + LPS + Si conditions. (E) Representative immunoblots. (F–H) Densitometry of PTBP1 (F), PKM1 (G), and PKM2/PKM1 (H). (I, J) MitoTracker and cytochrome c staining (I) and quantification of cytosolic cytochrome c (J). Scale bar: 20 µm. (K, L) Fluo-3/AM imaging (K) and Ca²⁺ quantification (L) Scale bar: 20 µm. Each symbol represents one independent cell-culture experiment (n = 3 biological replicates/group; technical replicates averaged per experiment). Data are mean ± SD. Statistics: one-way ANOVA with Tukey. A P < 0.001; B P = 0.45 (ns); C P < 0.001; D P < 0.001; F–H P < 0.001; J P < 0.001; L P < 0.001 (selected Tukey comparisons shown in the plots). ns, not significant; *P < 0.05, **P < 0.01, ***P < 0.001.

To assess whether these molecular alterations were accompanied by changes in mitochondrial injury-related phenotypes, we evaluated cytochrome c (Cyto C) release and calcium dynamics. Immunofluorescence revealed that LPS treatment induced robust cytosolic Cyto C accumulation, indicating mitochondrial outer membrane permeabilization. This response was significantly attenuated upon TCONS knockdown (Fig. 6I,J), consistent with preserved mitochondrial integrity. Additionally, intracellular calcium imaging using Fluo-3/AM indicated that LPS-triggered calcium overload was also prevented in both TCONS knockdown groups (Fig. 6K,L), consistent with improved calcium homeostasis.

Collectively, these results indicate that TCONS is associated with metabolic and mitochondrial vulnerability under inflammatory stress. Silencing TCONS rebalanced PKM isoform expression and attenuated mitochondrial injury-related phenotypes and calcium dysregulation.

TCONS knockdown is associated with attenuation of RV dysfunction and mitochondrial injury in vivo

Integrated functional and mechanistic validation shows that AAV9-cTnT–based cardiomyocyte-enriched knockdown of TCONS in PAH rats confers significant protection against acute RVF. As shown in the schematic (Fig. 7A), AAV9-cTnT-miR30-r-TCONS-ZsGreen was used to achieve stable myocardial TCONS suppression. Knockdown efficiency was confirmed in right ventricular tissues (Fig. 7B). Immunofluorescence analysis (Fig. S6) provided qualitative evidence of ZsGreen expression in RV tissue under the experimental conditions. In the same in vivo setting, Socs3 mRNA and pSTAT3 protein levels were not significantly altered following shTCONS treatment (Fig. S7), arguing against overt myocardial perturbation of this axis under the tested condition.

Fig. 7 — Cardiomyocyte-enriched AAV9-cTnT–mediated TCONS knockdown ameliorates right ventricular metabolic remodeling and preserves mitochondrial structure in vivo. (A) Workflow of AAV9-cTnT–mediated cardiac knockdown (PAH + sh-TCONS vs PAH + sh-NC) in monocrotaline-induced PAH rats followed by LPS challenge. (B) qRT–PCR of TCONS in RV confirming knockdown. (C) qRT–PCR of Ptbp1, Pkm1, Pkm2, and PKM2/PKM1 ratio. (D) Representative immunoblots of PTBP1, PKM1, PKM2 and GAPDH. (E) Densitometry of PTBP1, PKM1, PKM2 and PKM2/PKM1 ratio. (F) Representative TEM images of RV myocardium. Scale bar, 1 μm. (G) Quantification of mitochondrial area and mean mitochondrial size; subtype distribution (types I–IV) shown as percentages. (H) Representative confocal images of RV sections stained with DAPI, MitoTracker, and cytochrome c (Cyto C). Scale bar, 20 μm. (I) Quantification of cytosolic Cyto C fluorescence intensity.Each symbol represents one animal (B–E, I: n = 4/group; G: n = 5/group; technical measurements averaged per animal). Data are mean ± SD. Statistics: Two-tailed unpaired t-tests were used. Key comparisons: TCONS (B) P = 0.0499; PKM2/PKM1 mRNA (C) P = 0.0020; PTBP1 protein (E) P = 0.0144; PKM2 protein (E) P = 0.0081; PKM2/PKM1 protein (E) P = 0.0106; mitochondrial area (G) P = 0.0020; mitochondrial size (G) P = 0.0061; cytosolic Cyto C (I) P = 0.0048. Exact P values for all endpoints are provided in the Source Data. ns, not significant; *P < 0.05, **P < 0.01.

Both mRNA and protein analyses revealed that TCONS knockdown downregulated PTBP1 and PKM2, upregulated PKM1, and reduced the PKM2/PKM1 ratio (Fig. 7C–E), consistent with attenuation of a PKM2-dominant metabolic remodeling pattern. Transmission electron microscopy demonstrated marked improvement in mitochondrial morphology, as reflected by increased mitochondrial area, size, and normalization of mitochondrial subtypes (Fig. 7F,G). Furthermore, immunofluorescence analysis showed restored cytochrome c distribution, indicating preserved mitochondrial integrity (Fig. 7H,I).

Functionally, echocardiographic analysis (Fig. 8A) demonstrated that using a cardiomyocyte-enriched AAV9-cTnT strategy, TCONS knockdown was associated with attenuation of LPS-induced right ventricular dysfunction in PAH rats. Quantitative measurements revealed improvements in fractional area change (FAC), end-systolic and end-diastolic areas (RVA(s), RVA(d)), tricuspid annular plane systolic excursion (TAPSE), global longitudinal strain (GS), and free wall strain (FWS) in the TCONS knockdown group compared to controls (Fig. 8B–J). Representative M-mode and speckle-tracking images, as well as Doppler and strain imaging, further confirmed preserved right ventricular structure and contractile function following TCONS knockdown (Fig. 8E,H). Typical echocardiographic videos are provided in Supplementary Videos S1–S12. Importantly, Kaplan–Meier survival analysis showed a significant improvement in survival for PAH rats with TCONS knockdown after LPS challenge (Fig. 8K).

Furthermore, TCONS knockdown primarily regulates metabolic and mitochondrial adaptations, whereas established extracellular matrix remodeling and fibrosis appear less sensitive to this intervention. These findings underscore the multifactorial and irreversible nature of maladaptive cardiac remodeling in chronic PAH, highlighting the need for combinatorial or earlier interventions to effectively reverse fibrosis.

Discussion

Our study implicates lncRNA TCONS_00052110 (TCONS) as a candidate regulator of metabolic maladaptation in acute RVF, a decompensated stage associated with clinical deterioration in PAH. Mechanistically, TCONS physically associates with PTBP1 and is associated with prolonged half-life of PTBP1 protein, a higher PKM2/PKM1 ratio, and a PKM2-dominant metabolic state consistent with glycolysis-related remodeling, accompanied by mitochondrial injury and structural abnormalities. Using a cardiomyocyte-enriched AAV9-cTnT strategy^23,24, TCONS knockdown under inflammatory stress was associated with attenuation of acute RV dysfunction and mitochondrial injury, consistent with disruption of the TCONS–PTBP1–PKM2 axis.

These findings support the concept that lncRNA-driven metabolic dysregulation is one component underlying the inflammatory susceptibility of the right ventricle in PAH, thereby complementing and expanding the theory of the PTBP1/PKM2 axis in right ventricular metabolic remodeling and failure^12,25. Our data suggest that the TCONS–PTBP1–PKM2 axis acts as a candidate regulatory node facilitating glycolysis-related remodeling under inflammatory stress. The RV in the context of PAH is highly susceptible to inflammatory insults, readily transitioning from compensation to decompensation^2,3. This phenomenon is not solely due to the amplification of inflammatory signaling, but also reflects the intricate crosstalk between cardiac energy metabolism, the immune microenvironment, and cellular stress responses^11,26. Importantly, because we did not map TCONS induction across the full natural history of PAH (early adaptation to overt decompensation), our data primarily capture the acute inflammatory decompensation stage.

Building on our experimental findings, we further examined independent multi-omics datasets across species and tissues to contextualize these observations. These datasets included GO/KEGG pathway analyses derived from rodent proteomic and transcriptomic studies, as well as human transcriptomic datasets from PAH patient lung tissue and from right ventricular tissue representing compensated and decompensated RV states (Supplementary Figs S2 and S3). Although these datasets are analytically independent from our in vivo and in vitro experiments and do not directly quantify TCONS, the recurring enrichment of inflammatory, glycolytic, and mitochondrial pathways across species is concordant with the notion that metabolic remodeling and mitochondrial vulnerability may contribute to the inflammatory susceptibility of the RV in PAH.

Traditionally, RVF has long been regarded as a passive consequence of increased afterload secondary to left ventricular failure (LVF)²⁷. However, recent advances in both basic and clinical research have highlighted the unique pathophysiological role of the right ventricle in conditions such as PAH, leading to the identification of RV-specific molecular targets and therapeutic strategies^22,28,29. This paradigm shift has challenged the limitations of the traditional “left heart–centric” model. Given that the molecular mechanisms underlying right ventricular decompensation remain poorly understood, and conventional LV-targeted therapies demonstrate limited efficacy in RVF, there is an urgent need to establish pressure-overload RV failure models. Such models are essential to systematically elucidate RV-specific disease mechanisms and to pave the way for the development of innovative, RV-focused therapeutic approaches^25,30. Therefore, we propose that the inflammatory vulnerability of the RV in PAH may represent a “metabolic-inflammatory vulnerability window” in which chronic pressure overload creates a primed energetic state that is disproportionately destabilized by additional inflammatory stressors, with definable molecular mechanisms and clinical implications.

While most prior studies in the cardiovascular field have focused on the transcriptional regulation or ceRNA function of lncRNA^31–33, our work supports a model in which TCONS may modulate PTBP1 protein homeostasis by associating with reduced PTBP1 turnover post-transcriptionally, thereby influencing metabolic fate decisions in cardiomyocytes under inflammatory stress. This molecular mechanism positions TCONS as a potential upstream regulator of the inflammation-metabolism network and adds to, rather than replaces, the traditional protein-centric paradigm of metabolic regulation. The proposed TCONS–PTBP1 axis linked to reduced PTBP1 turnover further refines the “RBP–metabolic regulation” theory proposed by Caruso et al. and Du et al.^12,34. Moreover, we observed that TCONS upregulation is associated with amplification of PTBP1/PKM2-linked glycolytic remodeling in acute inflammatory and PAH two-hit models, highlighting the possibility that TCONS functions as a stress-amplifying hub and may represent a candidate target for future translational investigation.

Our study proposes that the TCONS-PTBP1-PKM2 axis may further modulate glycolysis-related remodeling during inflammatory stress and may serve as a candidate regulatory node influencing the transition from physiological adaptation to pathological decompensation in RV energetics. While glycolytic activation can be compensatory in some settings, our acute decompensation model suggests that a persistent PKM2-biased metabolic state consistent with glycolysis-related remodeling is associated with maladaptive RV remodeling. This finding not only qualifies and extends the traditional view of “beneficial glycolysis” but also aligns with the adverse outcomes of metabolic remodeling described by Riou et al. and Bornstein et al.^5,10,11. The novelty of this work lies in utilizing lncRNA as an entry point to connect the three core networks of inflammation, metabolism, and mitochondria, thereby suggesting a potential regulatory role in myocardial energy homeostasis and RV adaptation.

Cross-species analyses suggest partial conservation of genomic context at the TCONS locus, although direct conservation of transcript identity and function remains unverified in human tissue. Additionally, our multi-omics investigations corroborate the link between mitochondrial inflammatory metabolic disturbances and the pathogenesis of RVF. However, despite reanalyzing human lung and RV tissue datasets, we did not directly evaluate TCONS expression or function in human tissues, which limits inference regarding conservation and supports translational relevance in principle. Accordingly, our findings should be viewed as mechanistic evidence in preclinical models that warrants validation in human RV tissue and complementary RV stress models (e.g., SuHx, PAB) to strengthen generalizability. Our results are consistent with the molecular network of RV remodeling in PAH described by Shimauchi et al. and the comprehensive human cardiac metabolic studies by Bornstein et al.^11,25. We propose that energetic crisis and mitochondrial injury are consistent with a broader pathological theme rather than a phenomenon limited to experimental models, although this interpretation requires further validation.

With the clinical advancement of ASOs and other nucleic acid drugs, TCONS may represent a candidate molecular target for future precision-therapy-oriented research, pending further validation in human tissues. Prior studies by Lee et al. and Ma et al. have shown that antisense oligonucleotides targeting lncRNAs can effectively ameliorate cardiac pathology^35,36. Future research should focus on delineating the expression and regulation of TCONS in human populations, its interactions with other metabolic and mitochondrial pathways, and the synergistic effects of multi-target combination therapies^27,37. Collectively, our study provides a conceptual and experimental framework for RV-focused mechanistic studies and suggests that inflammation-triggered metabolic fragility may be therapeutically addressable at the post-transcriptional level.

Although TCONS knockdown effectively ameliorates metabolic remodeling-related abnormalities and attenuates mitochondrial injury, its impact on established right ventricular hypertrophy and fibrosis remains limited. This likely reflects the stage-specific nature of metabolic interventions, which can restore metabolic and mitochondrial function but are not expected to reverse fixed structural remodeling such as fibrosis. This dissociation between structural remodeling and functional recovery is consistent with both experimental and clinical observations by Shimauchi et al. and Guo et al.^14,25, and reflects the multifactorial and largely irreversible nature of chronic structural remodeling. Consequently, these findings suggest that the management of advanced RVF secondary to PAH should prioritize functional preservation and the delay of decompensation, rather than overemphasizing structural reversal—a concept relevant to timing and endpoint selection in future RVF-directed interventions.

In conclusion, our study supports TCONS as a candidate upstream regulator associated with inflammatory stress, metabolic remodeling, and mitochondrial injury in RVF. These findings provide a mechanistic framework that may inform future studies of lncRNA-targeted interventions aimed at preserving RV function and preventing acute RVF in PAH.

Limitations

This study has several limitations. First, our data mainly capture the stage of acute inflammatory decompensation in PAH rather than the full temporal course of RV adaptation and failure, because we did not assess the dynamics of TCONS induction across earlier and later disease stages. Second, although we reanalyzed human multi-omics datasets to contextualize our findings, we did not directly evaluate TCONS expression or function in human RV tissue, which limits inference on cross-species conservation and translational relevance. Third, at the TCONS/Socs3 overlapping locus, sequence-based analyses and RV Socs3 mRNA measurements argued against overt myocardial transcript-level suppression of Socs3 under the tested condition, but protein-level, signaling-level, extracardiac, and locus-level collateral effects cannot be fully excluded. Fourth, the current design does not formally dissociate acute hemodynamic afterload from myocardial metabolic and mitochondrial injury programs, and these abnormalities should therefore be interpreted as biologically relevant associated components of acute RV decompensation rather than isolated primary drivers. Fifth, the evidence supporting TCONS localization and interaction with PTBP1 remains supportive rather than fully orthogonal, because we did not perform nuclear–cytoplasmic fractionation, additional RNA-FISH probe-level controls, purified-component binding assays, or motif-disrupting rescue experiments; likewise, although our in vitro data support reduced PTBP1 turnover, we did not perform proteasome inhibition or ubiquitination assays to define the precise degradation pathway. Sixth, our metabolic and mitochondrial conclusions are limited by the absence of direct glycolytic flux measurements and by the lack of post-knockdown RV bioenergetic assays, such as high-resolution respirometry or ATP quantification in treated hearts, such that the present data support glycolysis-related remodeling and attenuated mitochondrial injury, but not definitive restoration of energetic output. Finally, although TCONS knockdown improved metabolic and mitochondrial phenotypes, it had limited effects on established RV hypertrophy and fibrosis, and we did not quantify AAV9 transduction efficiency, perform detailed extracardiac tissue profiling, or include PTBP1/PKM2 rescue experiments, all of which would further strengthen causal and tissue-specific interpretation.

Methods

Animal model of acute RVF in PAH

All animal procedures were approved by the Institutional Animal Care and Use Committee of Xiangya Hospital, Central South University (Approval No. 201303311) and conducted in accordance with NIH and ARRIVE guidelines. Adult male Sprague–Dawley rats (250–300 g) were housed under standard conditions with ad libitum access to food and water.

Pulmonary arterial hypertension (PAH) was induced by intraperitoneal injection of monocrotaline (MCT, 60 mg/kg). Control animals received equal volumes of saline. On day 28 after injection, transthoracic echocardiography was used to confirm PAH establishment (defined as systolic pulmonary arterial pressure [sPAP] > 60 mmHg). Only confirmed PAH rats were used for subsequent experiments.

To simulate acute right ventricular failure (RVF) following mild systemic inflammatory stress, lipopolysaccharide (LPS, 1 mg/kg) was injected intraperitoneally into PAH and controls on day 28. This protocol mimics clinical inflammatory triggers (e.g., infection or surgical stress) that may precipitate acute RVF¹⁵. The selected LPS dose (1 mg/kg) has been validated in our previous work to induce a transient systemic inflammatory response without direct cardiotoxicity in PAH or control rats¹⁶. Echocardiography was performed before and 2 h after LPS. Detailed animal procedures are provided in Supplementary Methods (S1).

Echocardiography

Right ventricular (RV) function was assessed by transthoracic echocardiography at predefined experimental time points, including the PAH stage and after AAV9-cTnT–based TCONS modulation. Parameters included tricuspid annular plane systolic excursion (TAPSE), right ventricular fractional area change (FAC), strain-based indices, RV dimensions, and heart rate. All measurements were performed by a trained operator blinded to group allocation according to standardized guidelines. Detailed acquisition parameters, calibration procedures, and reproducibility assessments are provided in Supplementary Methods (S2).

RNA isolation and quantitative real-time PCR (qRT-PCR)

Total RNA was extracted from rat RV tissues and cultured cells, and lncRNA and mRNA expression levels were quantified by quantitative real-time PCR. Relative expression was calculated using the 2 ^∧ − ΔΔCt method. Primer sequences, reaction conditions, replication details, and the in silico assessment of assay-level specificity for the current TCONS_00052110 qRT-PCR primer pair are provided in the Supplementary Methods and Supplementary Tables (S3, S4; Tables S1 and S7).

Protein expression analysis

Protein expression levels were assessed by immunoblotting and immunofluorescence as indicated. Detailed antibody information and experimental procedures are provided in the Supplementary Methods (S5).

AAV-mediated TCONS modulation in vivo

AAV9 vectors driven by a cardiomyocyte-enriched cTnT promoter were used to modulate TCONS expression in vivo. Adult rats were randomly assigned to experimental or control groups and received a single tail-vein injection of TCONS shRNA or control vectors at the indicated dose. PAH was induced four weeks after AAV administration, followed by an additional stabilization period prior to LPS challenge. In the same in vivo setting, RV Socs3 mRNA and pSTAT3 levels were assessed at the experimental endpoint to examine whether shTCONS treatment was associated with overt perturbation of the myocardial Socs3–STAT3 axis under the tested condition. In addition, sequence-liability assessment of the TCONS-targeting RNAi reagents at the TCONS/Socs3 locus was performed in silico. Detailed vector construction, titration, endpoint validation, and RNAi liability assessment procedures are provided in Supplementary Methods (S6–S8; Table S2).

Histology and morphological analysis

Right ventricular remodeling was evaluated by histological staining, and mitochondrial ultrastructure was assessed by transmission electron microscopy. Quantitative analyses were performed in a blinded manner. Detailed staining procedures, morphometric criteria, and mitochondrial classification methods are described in Supplementary Methods (S9, S10).

Mitochondrial functional assessment

Mitochondrial oxidative phosphorylation capacity in RV tissue was evaluated using high-resolution respirometry under defined substrate–inhibitor conditions. Detailed experimental protocols and quality control procedures are provided in Supplementary Methods (S11).

Cell culture and in vitro treatments

H9C2 rat cardiomyoblasts were used for mechanistic in vitro studies. Cells were subjected to hypertrophic and inflammatory stimulation, followed by TCONS knockdown or overexpression as indicated. Detailed culture conditions, treatments, and transfection procedures are provided in Supplementary Methods (S12; Tables S3 and S4).

Immunofluorescence and functional cellular assays

Immunofluorescence staining, intracellular reactive oxygen species detection, cytochrome c localization, intracellular calcium measurement, RNA-FISH, RNA pulldown assays, and cycloheximide chase experiments were performed to investigate mitochondrial dysfunction and PTBP1 stability. Detailed experimental procedures, including probe and primer information for RNA-based assays, are provided in Supplementary Methods (S13–S21) and Supplementary Table S5.

Clinical omics dataset analysis

Publicly available transcriptomic datasets were retrieved from NCBI GEO and reprocessed de novo for differential expression and pathway enrichment analyses. Group definitions followed the original studies. Detailed bioinformatic pipelines and scripts are provided in Supplementary Methods (S22).

Statistical analysis

Data are presented as mean ± standard deviation (SD). Comparisons between two groups were performed using unpaired t-tests. One-way ANOVA was used for single-factor comparisons, and two-way ANOVA was applied for analyses involving two independent factors, followed by Tukey’s multiple-comparisons test. Categorical variables were analyzed using chi-square tests. Biological replicate numbers are reported in the figure legends, and technical replicates were averaged within each biological replicate. Detailed statistical considerations are provided in Supplementary Methods (S23).

Supplementary Information

Supplementary Information 1.^{(2.8MB, pdf)}

Supplementary Video 1.^{(22.6MB, avi)}

Supplementary Video 2.^{(2.7MB, avi)}

Supplementary Video 3.^{(2.7MB, avi)}

Supplementary Video 4.^{(7.2MB, avi)}

Supplementary Video 5.^{(25.2MB, avi)}

Supplementary Video 6.^{(38.8MB, avi)}

Supplementary Video 7.^{(42.2MB, avi)}

Supplementary Video 8.^{(16.1MB, avi)}

Supplementary Video 9.^{(26.8MB, avi)}

Supplementary Video 10.^{(13.6MB, avi)}

Supplementary Video 11.^{(17.2MB, avi)}

Supplementary Video 12.^{(3.1MB, avi)}

Author contributions

Xiaowei Gao: Formal analysis; Visualization; Writing—original draft; Writing—review & editing. Yue Yang: Investigation. Lizhe Guo: Formal analysis. Lu Wang: Formal analysis. Qian Li: Visualization. Gang Qin: Writing—review & editing. Hui Luo: Conceptualization; Methodology; Supervision. Yanan Cao: Conceptualization; Methodology; Interpretation; Writing—review & editing; Supervision. E Wang: Conceptualization; Methodology; Interpretation; Writing—review & editing; Supervision. All authors reviewed and approved the final manuscript.

Funding

National Natural Science Foundation of China (NSFC, General Program, Grant Nos. 81873508 and 81800058, to Prof. E. Wang); National Natural Science Foundation of China (NSFC, Young Scientists Fund, Grant Nos. 82000388 and 82200324, to Hui Luo and Yanan Cao); Natural Science Foundation of Hunan Province (Grant Nos. 2020JJ4900 and 2023JJ40926, to Hui Luo and Yanan Cao).

Data availability

All data generated or analyzed in this study are included in this article and its supplementary information files. Uncropped Western blot images are provided in the supplementary information. Additional raw data and analysis codes are available from the corresponding author upon reasonable request. Publicly available transcriptomic datasets were obtained from the Gene Expression Omnibus (GEO) under accession numbers GSE117261 (PAH and control lung tissue transcriptomes) and GSE240921 (human right ventricular tissue transcriptomes of adaptive vs maladaptive remodeling). This study did not recruit human participants and did not collect new human samples; only de-identified, publicly available datasets were analyzed and no additional ethical approval was required.

Declarations

Competing interests

The authors declare no competing interests.

Footnotes

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Contributor Information

Yanan Cao, Email: 893085967@qq.com.

E. Wang, Email: ewang324@csu.edu.cn

References

1.Garcia, M. V. F., Nanah, A. & Caruso, P. Acute decompensated pulmonary hypertension outcomes in pulmonary arterial hypertension patients: Systematic review and meta-analysis of proportions. Heart Lung70, 57–64. 10.1016/j.hrtlng.2024.11.009 (2025). [DOI] [PubMed]
2.Giannakoulas, G. et al. Acute right ventricular failure: Pathophysiology, aetiology, assessment, and management. Eur. Heart J.10.1093/eurheartj/ehaf215 (2025). [DOI] [PubMed]
3.Olsson, K. M. et al. Right heart function during and after pregnancy in women with pulmonary arterial hypertension. Eur. Respir. J.10.1183/13993003.00179-2024 (2024). [DOI] [PMC free article] [PubMed]
4.Vasudeva, R., Challa, A., Tuck, N., Pothuru, S. & Vindhyal, M. Hospital outcomes for patients with pulmonary arterial hypertension in sepsis and septic shock. BMC Pulm. Med.22, 374. 10.1186/s12890-022-02145-1 (2022). [DOI] [PMC free article] [PubMed]
5.Riou, M., Enache, I., Sauer, F., Charles, A. L. & Geny, B. Targeting mitochondrial metabolic dysfunction in pulmonary hypertension: Toward new therapeutic approaches?. Int. J. Mol. Sci.10.3390/ijms24119572 (2023). [DOI] [PMC free article] [PubMed]
6.Esparza-Moltó, P. B. & Cuezva, J. M. Reprogramming oxidative phosphorylation in cancer: A role for RNA-binding proteins. Antioxid. Redox Signal.33, 927–945. 10.1089/ars.2019.7988 (2020). [DOI] [PubMed]
7.Zhou, R., Yazdi, A. S., Menu, P. & Tschopp, J. A role for mitochondria in NLRP3 inflammasome activation. Nature469, 221–225. 10.1038/nature09663 (2011). [DOI] [PubMed]
8.Zorov, D. B., Juhaszova, M. & Sollott, S. J. Mitochondrial reactive oxygen species (ROS) and ROS-induced ROS release. Physiol. Rev.94, 909–950. 10.1152/physrev.00026.2013 (2014). [DOI] [PMC free article] [PubMed]
9.Zecchini, V. et al. Fumarate induces vesicular release of mtDNA to drive innate immunity. Nature615, 499–506. 10.1038/s41586-023-05770-w (2023). [DOI] [PMC free article] [PubMed]
10.Xu, W., Janocha, A. J. & Erzurum, S. C. Metabolism in pulmonary hypertension. Annu. Rev. Physiol.83, 551–576. 10.1146/annurev-physiol-031620-123956 (2021). [DOI] [PMC free article] [PubMed]
11.Bornstein, M. R., Tian, R. & Arany, Z. Human cardiac metabolism. Cell Metab.36, 1456–1481. 10.1016/j.cmet.2024.06.003 (2024). [DOI] [PMC free article] [PubMed]
12.Caruso, P. et al. Identification of MicroRNA-124 as a major regulator of enhanced endothelial cell glycolysis in pulmonary arterial hypertension via PTBP1 (polypyrimidine tract binding protein) and pyruvate kinase M2. Circulation136, 2451–2467. 10.1161/circulationaha.117.028034 (2017). [DOI] [PMC free article] [PubMed]
13.Glisovic, T., Bachorik, J. L., Yong, J. & Dreyfuss, G. RNA-binding proteins and post-transcriptional gene regulation. FEBS Lett.582, 1977–1986. 10.1016/j.febslet.2008.03.004 (2008). [DOI] [PMC free article] [PubMed]
14.Guo, L. et al. M-type pyruvate kinase 2 (PKM2) tetramerization alleviates the progression of right ventricle failure by regulating oxidative stress and mitochondrial dynamics. J. Transl. Med.21, 888. 10.1186/s12967-023-04780-6 (2023). [DOI] [PMC free article] [PubMed]
15.Guo, L. et al. Regulation of the immune microenvironment by an NLRP3 inhibitor contributes to attenuation of acute right ventricular failure in rats with pulmonary arterial hypertension. J. Inflamm. Res.14, 5699–5711. 10.2147/jir.S336964 (2021). [DOI] [PMC free article] [PubMed]
16.Zhang, J. et al. Lipopolysaccharide acutely suppresses right-ventricular strain in rats with pulmonary artery hypertension. Pulm. Circ.8, 2045893217744504. 10.1177/2045893217744504 (2018). [DOI] [PMC free article] [PubMed]
17.Yang, Y. et al. Long non-coding RNA expression profiling in the lungs of pulmonary arterial hypertension rats with acute inflammation. Pulm. Circ.9, 2045894019879393. 10.1177/2045894019879393 (2019). [DOI] [PMC free article] [PubMed]
18.Cao, Y. et al. Analyses of long non-coding RNA and mRNA profiles in right ventricle myocardium of acute right heart failure in pulmonary arterial hypertension rats. Biomed. Pharmacother.106, 1108–1115. 10.1016/j.biopha.2018.07.057 (2018). [DOI] [PubMed]
19.Zahid, K. R., Raza, U., Chen, J., Raj, U. J. & Gou, D. Pathobiology of pulmonary artery hypertension: Role of long non-coding RNAs. Cardiovasc. Res.116, 1937–1947. 10.1093/cvr/cvaa050 (2020). [DOI] [PubMed]
20.Han, D., Gao, Q. & Cao, F. Long noncoding RNAs (LncRNAs) - The dawning of a new treatment for cardiac hypertrophy and heart failure. Biochim. Biophys. Acta (BBA) Mol. Basis Dis.1863, 2078–2084. 10.1016/j.bbadis.2017.02.024 (2017). [DOI] [PubMed]
21.Yang, Y. et al. Plasma MIR-212-3p as a biomarker for acute right heart failure with pulmonary artery hypertension. Ann. Transl. Med.8, 1571. 10.21037/atm-20-1653a (2020). [DOI] [PMC free article] [PubMed]
22.Klein, M. et al. Combined tyrosine and serine/threonine kinase inhibition by sorafenib prevents progression of experimental pulmonary hypertension and myocardial remodeling. Circulation118, 2081–2090. 10.1161/circulationaha.108.779751 (2008). [DOI] [PubMed]
23.Prasad, K. M., Xu, Y., Yang, Z., Acton, S. T. & French, B. A. Robust cardiomyocyte-specific gene expression following systemic injection of AAV: In vivo gene delivery follows a Poisson distribution. Gene Ther.18, 43–52. 10.1038/gt.2010.105 (2011). [DOI] [PMC free article] [PubMed]
24.Werfel, S. et al. Rapid and highly efficient inducible cardiac gene knockout in adult mice using AAV-mediated expression of Cre recombinase. Cardiovasc. Res.104, 15–23. 10.1093/cvr/cvu174 (2014). [DOI] [PubMed]
25.Shimauchi, T. et al. PARP1-PKM2 axis mediates right ventricular failure associated with pulmonary arterial hypertension. JACC. Basic Transl. Sci.7, 384–403. 10.1016/j.jacbts.2022.01.005 (2022). [DOI] [PMC free article] [PubMed]
26.Savale, L. et al. Acute decompensated pulmonary hypertension. Eur. Respir. Rev.26, 170092. 10.1183/16000617.0092-2017 (2017). [DOI] [PMC free article] [PubMed]
27.Arici, M. et al. Selective SERCA2a activator as a candidate for chronic heart failure therapy. J. Transl. Med.22, 77. 10.1186/s12967-024-04874-9 (2024). [DOI] [PMC free article] [PubMed]
28.Khassafi, F. et al. Transcriptional profiling unveils molecular subgroups of adaptive and maladaptive right ventricular remodeling in pulmonary hypertension. Nat. Cardiovasc. Res.2, 917–936. 10.1038/s44161-023-00338-3 (2023). [DOI] [PMC free article] [PubMed]
29.Lajoie, A. C. et al. Combination therapy versus monotherapy for pulmonary arterial hypertension: A meta-analysis. Lancet Respir. Med.4, 291–305. 10.1016/s2213-2600(16)00027-8 (2016). [DOI] [PubMed]
30.Reddy, S. & Bernstein, D. Molecular mechanisms of right ventricular failure. Circulation132, 1734–1742. 10.1161/circulationaha.114.012975 (2015). [DOI] [PMC free article] [PubMed]
31.Guttman, M. & Rinn, J. L. Modular regulatory principles of large non-coding RNAs. Nature482, 339–346. 10.1038/nature10887 (2012). [DOI] [PMC free article] [PubMed]
32.Li, J. et al. Long noncoding RNA NRON contributes to HIV-1 latency by specifically inducing tat protein degradation. Nat. Commun.7, 11730. 10.1038/ncomms11730 (2016). [DOI] [PMC free article] [PubMed]
33.Mongelli, A., Martelli, F., Farsetti, A. & Gaetano, C. The dark that matters: Long non-coding RNAs as master regulators of cellular metabolism in non-communicable diseases. Front. Physiol.10, 369. 10.3389/fphys.2019.00369 (2019). [DOI] [PMC free article] [PubMed]
34.Du, H. et al. LncRNA DCRT protects against dilated cardiomyopathy by preventing NDUFS2 alternative splicing by binding to PTBP1. Circulation150, 1030–1049. 10.1161/circulationaha.123.067861 (2024). [DOI] [PubMed]
35.Lee, R. G., Crosby, J., Baker, B. F., Graham, M. J. & Crooke, R. M. Antisense technology: An emerging platform for cardiovascular disease therapeutics. J. Cardiovasc. Transl. Res.6, 969–980. 10.1007/s12265-013-9495-7 (2013). [DOI] [PMC free article] [PubMed]
36.Ma, C. et al. Super enhancer-associated circular RNA-CircKrt4 regulates hypoxic pulmonary artery endothelial cell dysfunction in mice. Arterioscler. Thromb. Vasc. Biol.43, 1179–1198. 10.1161/atvbaha.122.318842 (2023). [DOI] [PubMed]
37.Chiao, Y. A. et al. Late-life restoration of mitochondrial function reverses cardiac dysfunction in old mice. Elife10.7554/eLife.55513 (2020). [DOI] [PMC free article] [PubMed]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

Supplementary Information 1.^{(2.8MB, pdf)}

Supplementary Video 1.^{(22.6MB, avi)}

Supplementary Video 2.^{(2.7MB, avi)}

Supplementary Video 3.^{(2.7MB, avi)}

Supplementary Video 4.^{(7.2MB, avi)}

Supplementary Video 5.^{(25.2MB, avi)}

Supplementary Video 6.^{(38.8MB, avi)}

Supplementary Video 7.^{(42.2MB, avi)}

Supplementary Video 8.^{(16.1MB, avi)}

Supplementary Video 9.^{(26.8MB, avi)}

Supplementary Video 10.^{(13.6MB, avi)}

Supplementary Video 11.^{(17.2MB, avi)}

Supplementary Video 12.^{(3.1MB, avi)}

Data Availability Statement

[CR1] 1.Garcia, M. V. F., Nanah, A. & Caruso, P. Acute decompensated pulmonary hypertension outcomes in pulmonary arterial hypertension patients: Systematic review and meta-analysis of proportions. Heart Lung70, 57–64. 10.1016/j.hrtlng.2024.11.009 (2025). [DOI] [PubMed]

[CR2] 2.Giannakoulas, G. et al. Acute right ventricular failure: Pathophysiology, aetiology, assessment, and management. Eur. Heart J.10.1093/eurheartj/ehaf215 (2025). [DOI] [PubMed]

[CR3] 3.Olsson, K. M. et al. Right heart function during and after pregnancy in women with pulmonary arterial hypertension. Eur. Respir. J.10.1183/13993003.00179-2024 (2024). [DOI] [PMC free article] [PubMed]

[CR4] 4.Vasudeva, R., Challa, A., Tuck, N., Pothuru, S. & Vindhyal, M. Hospital outcomes for patients with pulmonary arterial hypertension in sepsis and septic shock. BMC Pulm. Med.22, 374. 10.1186/s12890-022-02145-1 (2022). [DOI] [PMC free article] [PubMed]

[CR5] 5.Riou, M., Enache, I., Sauer, F., Charles, A. L. & Geny, B. Targeting mitochondrial metabolic dysfunction in pulmonary hypertension: Toward new therapeutic approaches?. Int. J. Mol. Sci.10.3390/ijms24119572 (2023). [DOI] [PMC free article] [PubMed]

[CR6] 6.Esparza-Moltó, P. B. & Cuezva, J. M. Reprogramming oxidative phosphorylation in cancer: A role for RNA-binding proteins. Antioxid. Redox Signal.33, 927–945. 10.1089/ars.2019.7988 (2020). [DOI] [PubMed]

[CR7] 7.Zhou, R., Yazdi, A. S., Menu, P. & Tschopp, J. A role for mitochondria in NLRP3 inflammasome activation. Nature469, 221–225. 10.1038/nature09663 (2011). [DOI] [PubMed]

[CR8] 8.Zorov, D. B., Juhaszova, M. & Sollott, S. J. Mitochondrial reactive oxygen species (ROS) and ROS-induced ROS release. Physiol. Rev.94, 909–950. 10.1152/physrev.00026.2013 (2014). [DOI] [PMC free article] [PubMed]

[CR9] 9.Zecchini, V. et al. Fumarate induces vesicular release of mtDNA to drive innate immunity. Nature615, 499–506. 10.1038/s41586-023-05770-w (2023). [DOI] [PMC free article] [PubMed]

[CR10] 10.Xu, W., Janocha, A. J. & Erzurum, S. C. Metabolism in pulmonary hypertension. Annu. Rev. Physiol.83, 551–576. 10.1146/annurev-physiol-031620-123956 (2021). [DOI] [PMC free article] [PubMed]

[CR11] 11.Bornstein, M. R., Tian, R. & Arany, Z. Human cardiac metabolism. Cell Metab.36, 1456–1481. 10.1016/j.cmet.2024.06.003 (2024). [DOI] [PMC free article] [PubMed]

[CR12] 12.Caruso, P. et al. Identification of MicroRNA-124 as a major regulator of enhanced endothelial cell glycolysis in pulmonary arterial hypertension via PTBP1 (polypyrimidine tract binding protein) and pyruvate kinase M2. Circulation136, 2451–2467. 10.1161/circulationaha.117.028034 (2017). [DOI] [PMC free article] [PubMed]

[CR13] 13.Glisovic, T., Bachorik, J. L., Yong, J. & Dreyfuss, G. RNA-binding proteins and post-transcriptional gene regulation. FEBS Lett.582, 1977–1986. 10.1016/j.febslet.2008.03.004 (2008). [DOI] [PMC free article] [PubMed]

[CR14] 14.Guo, L. et al. M-type pyruvate kinase 2 (PKM2) tetramerization alleviates the progression of right ventricle failure by regulating oxidative stress and mitochondrial dynamics. J. Transl. Med.21, 888. 10.1186/s12967-023-04780-6 (2023). [DOI] [PMC free article] [PubMed]

[CR15] 15.Guo, L. et al. Regulation of the immune microenvironment by an NLRP3 inhibitor contributes to attenuation of acute right ventricular failure in rats with pulmonary arterial hypertension. J. Inflamm. Res.14, 5699–5711. 10.2147/jir.S336964 (2021). [DOI] [PMC free article] [PubMed]

[CR16] 16.Zhang, J. et al. Lipopolysaccharide acutely suppresses right-ventricular strain in rats with pulmonary artery hypertension. Pulm. Circ.8, 2045893217744504. 10.1177/2045893217744504 (2018). [DOI] [PMC free article] [PubMed]

[CR17] 17.Yang, Y. et al. Long non-coding RNA expression profiling in the lungs of pulmonary arterial hypertension rats with acute inflammation. Pulm. Circ.9, 2045894019879393. 10.1177/2045894019879393 (2019). [DOI] [PMC free article] [PubMed]

[CR18] 18.Cao, Y. et al. Analyses of long non-coding RNA and mRNA profiles in right ventricle myocardium of acute right heart failure in pulmonary arterial hypertension rats. Biomed. Pharmacother.106, 1108–1115. 10.1016/j.biopha.2018.07.057 (2018). [DOI] [PubMed]

[CR19] 19.Zahid, K. R., Raza, U., Chen, J., Raj, U. J. & Gou, D. Pathobiology of pulmonary artery hypertension: Role of long non-coding RNAs. Cardiovasc. Res.116, 1937–1947. 10.1093/cvr/cvaa050 (2020). [DOI] [PubMed]

[CR20] 20.Han, D., Gao, Q. & Cao, F. Long noncoding RNAs (LncRNAs) - The dawning of a new treatment for cardiac hypertrophy and heart failure. Biochim. Biophys. Acta (BBA) Mol. Basis Dis.1863, 2078–2084. 10.1016/j.bbadis.2017.02.024 (2017). [DOI] [PubMed]

[CR21] 21.Yang, Y. et al. Plasma MIR-212-3p as a biomarker for acute right heart failure with pulmonary artery hypertension. Ann. Transl. Med.8, 1571. 10.21037/atm-20-1653a (2020). [DOI] [PMC free article] [PubMed]

[CR22] 22.Klein, M. et al. Combined tyrosine and serine/threonine kinase inhibition by sorafenib prevents progression of experimental pulmonary hypertension and myocardial remodeling. Circulation118, 2081–2090. 10.1161/circulationaha.108.779751 (2008). [DOI] [PubMed]

[CR23] 23.Prasad, K. M., Xu, Y., Yang, Z., Acton, S. T. & French, B. A. Robust cardiomyocyte-specific gene expression following systemic injection of AAV: In vivo gene delivery follows a Poisson distribution. Gene Ther.18, 43–52. 10.1038/gt.2010.105 (2011). [DOI] [PMC free article] [PubMed]

[CR24] 24.Werfel, S. et al. Rapid and highly efficient inducible cardiac gene knockout in adult mice using AAV-mediated expression of Cre recombinase. Cardiovasc. Res.104, 15–23. 10.1093/cvr/cvu174 (2014). [DOI] [PubMed]

[CR25] 25.Shimauchi, T. et al. PARP1-PKM2 axis mediates right ventricular failure associated with pulmonary arterial hypertension. JACC. Basic Transl. Sci.7, 384–403. 10.1016/j.jacbts.2022.01.005 (2022). [DOI] [PMC free article] [PubMed]

[CR26] 26.Savale, L. et al. Acute decompensated pulmonary hypertension. Eur. Respir. Rev.26, 170092. 10.1183/16000617.0092-2017 (2017). [DOI] [PMC free article] [PubMed]

[CR27] 27.Arici, M. et al. Selective SERCA2a activator as a candidate for chronic heart failure therapy. J. Transl. Med.22, 77. 10.1186/s12967-024-04874-9 (2024). [DOI] [PMC free article] [PubMed]

[CR28] 28.Khassafi, F. et al. Transcriptional profiling unveils molecular subgroups of adaptive and maladaptive right ventricular remodeling in pulmonary hypertension. Nat. Cardiovasc. Res.2, 917–936. 10.1038/s44161-023-00338-3 (2023). [DOI] [PMC free article] [PubMed]

[CR29] 29.Lajoie, A. C. et al. Combination therapy versus monotherapy for pulmonary arterial hypertension: A meta-analysis. Lancet Respir. Med.4, 291–305. 10.1016/s2213-2600(16)00027-8 (2016). [DOI] [PubMed]

[CR30] 30.Reddy, S. & Bernstein, D. Molecular mechanisms of right ventricular failure. Circulation132, 1734–1742. 10.1161/circulationaha.114.012975 (2015). [DOI] [PMC free article] [PubMed]

[CR31] 31.Guttman, M. & Rinn, J. L. Modular regulatory principles of large non-coding RNAs. Nature482, 339–346. 10.1038/nature10887 (2012). [DOI] [PMC free article] [PubMed]

[CR32] 32.Li, J. et al. Long noncoding RNA NRON contributes to HIV-1 latency by specifically inducing tat protein degradation. Nat. Commun.7, 11730. 10.1038/ncomms11730 (2016). [DOI] [PMC free article] [PubMed]

[CR33] 33.Mongelli, A., Martelli, F., Farsetti, A. & Gaetano, C. The dark that matters: Long non-coding RNAs as master regulators of cellular metabolism in non-communicable diseases. Front. Physiol.10, 369. 10.3389/fphys.2019.00369 (2019). [DOI] [PMC free article] [PubMed]

[CR34] 34.Du, H. et al. LncRNA DCRT protects against dilated cardiomyopathy by preventing NDUFS2 alternative splicing by binding to PTBP1. Circulation150, 1030–1049. 10.1161/circulationaha.123.067861 (2024). [DOI] [PubMed]

[CR35] 35.Lee, R. G., Crosby, J., Baker, B. F., Graham, M. J. & Crooke, R. M. Antisense technology: An emerging platform for cardiovascular disease therapeutics. J. Cardiovasc. Transl. Res.6, 969–980. 10.1007/s12265-013-9495-7 (2013). [DOI] [PMC free article] [PubMed]

[CR36] 36.Ma, C. et al. Super enhancer-associated circular RNA-CircKrt4 regulates hypoxic pulmonary artery endothelial cell dysfunction in mice. Arterioscler. Thromb. Vasc. Biol.43, 1179–1198. 10.1161/atvbaha.122.318842 (2023). [DOI] [PubMed]

[CR37] 37.Chiao, Y. A. et al. Late-life restoration of mitochondrial function reverses cardiac dysfunction in old mice. Elife10.7554/eLife.55513 (2020). [DOI] [PMC free article] [PubMed]

PERMALINK

TCONS_00052110 is associated with prolonged PTBP1 half-life and mitochondrial injury in PAH-associated right ventricular stress

Xiaowei Gao

Yue Yang

Lizhe Guo

Lu Wang

Li Qian

Gang Qin

Hui Luo

Yanan Cao

E Wang

Abstract

Supplementary Information

Introduction

Results

Inflammatory stress precipitates acute right ventricular failure in PAH accompanied by glycolysis-related remodeling and mitochondrial injury

Fig. 1.

Fig. 2.

LncRNA TCONS_00052110 is induced in RVF rats and is characterized as a novel full-length noncoding transcript with post-transcriptional regulatory potential

Fig. 3.

Inflammatory two-hit stress drives oxidative and mitochondrial injury in hypertrophic cardiomyocytes

Fig. 4.

TCONS is associated with PTBP1 accumulation and reduced PTBP1 protein turnover in vitro

Fig. 5.

TCONS knockdown attenuates mitochondrial injury and disrupts the PTBP1–PKM2 axis

Fig. 6.

TCONS knockdown is associated with attenuation of RV dysfunction and mitochondrial injury in vivo

Fig. 7.

Fig. 8.

Discussion

Limitations

Methods

Animal model of acute RVF in PAH

Echocardiography

RNA isolation and quantitative real-time PCR (qRT-PCR)

Protein expression analysis

AAV-mediated TCONS modulation in vivo

Histology and morphological analysis

Mitochondrial functional assessment

Cell culture and in vitro treatments

Immunofluorescence and functional cellular assays

Clinical omics dataset analysis

Statistical analysis

Supplementary Information

Author contributions

Funding

Data availability

Declarations

Competing interests

Footnotes

Contributor Information

References

Associated Data

Supplementary Materials

Data Availability Statement

ACTIONS

PERMALINK

RESOURCES

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