Non-coding RNA transcriptome analysis of circulating human monocytes reveals subpopulation-enriched transcripts and selective isoforms differentially expressed during acute myocardial infarction

Background Prolonged delay in seeking treatment by patients with acute myocardial infarction (AMI) remains a significant therapeutic gap. Previous mass-media campaigns or brief educational interventions were largely ineffective in shortening pre-hospital patient delay. The brevity and didactic nature of these interventions are inadequate to address complex decision-making encountered by patients with AMI. Purpose This randomized controlled trial examined the effects of a theory-driven modelling-based narrative intervention on AMI knowledge, care-seeking intention as indicated by attitudes and beliefs regarding care-seeking for AMI, prehospital delay time and use of ambulance among AMI survivors. Methods A total of 285 of patients with history of at least one AMI were randomized 1:1 to modelling-based narrative intervention (n=144) and control with didactic education (n=141). The modelling-based narrative intervention comprised 4 weekly group-based interactive sessions to engage participants in mental rehearsals of the perceptual-cognitive processes in recognizing and responding to AMI symptoms through a virtual heart attack experience, then followed by a booster session one month later The control group received a 4-weekly education on factual information about AMI with traditional didactic education approach. Study endpoints included AMI knowledge, attitudes and belief about care-seeking for AMI to be measured at baseline, (T0), 3 months (T1), 12 months (T2) and 24 months (T3) after the intervention. Prehospital delay time and use of ambulance for an AMI attack were tracked since patients' enrolment until T3 endpoint of the study. Results Generalized estimating equation analysis indicated that the participants in the intervention group had significantly greater improvement in attitudes and beliefs about care seeking for AMI symptoms at all endpoints than those in the control group (Table 1). There were no between-group differences in the change of AMI knowledge score over the baseline and 3- or 12-month endpoints, such difference became significant at 24-month endpoint. A total of 78 participants, including 38 (26.4%) in the intervention group and 40 (28.4%) in the control groups, had experienced AMI symptoms and admitted to an emergency department during the follow-up period. The participants in the intervention group showed significant reduction in prehospital delay time than the control group (p=0.031). There was no between-group difference on use of ambulance (p=0.422) for those participants who had symptom attack during the follow-up period. Conclusion This study demonstrated favorable effects in shortening prehospital time delay and promoting positive attitudes and beliefs regarding care-seeking for AMI symptoms among AMI survivors. A virtual heart attack experience helped support them through a cognitive-perceptual processes in recognizing and interpreting AMI symptoms. Funding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Early Career Scheme, Research Grants Council, Hong Kong

Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): National Natural Science Foundation of China (NSFC). Introduction Numerous epidemiologic studies have confirmed that Standard Modifiable Cardiovascular Risk Factors (SMuRFs) are independent risk factors for coronary heart disease (CHD) and are recommended by guidelines for the primary and secondary prevention of CHD. However, it is interesting to find that the increasing proportion of individuals without SMuRFs in CHD and acute myocardial infarction (AMI). Moreover, accumulating clinical trials have reported the inverse association between the number of risk factors and crude mortality in AMI. It is unknown whether this paradoxical phenomenon can be seen in critical ill patients. Purpose To analyze and validate whether the number of risk factors contradicts the mortality outcome in patients with AMI in the MIMIC-III (Medical Information Mart for Intensive Care III) database. Methods A total of 2484 AMI patients (62.8% male, mean age 71.2 [59.3;80.8] years) admitting to cardiac care unit for the first time from the MIMIC-III database were enrolled. Risk factors were defined as hypertension, diabetes,dyslipidemia, and smoking history. They were grouped according to the number of risk factors. Main outcome was mortality at 2-year following hospitalization. Kaplan-Meier curve and Cox proportional hazards models adjusting for baseline clinical characteristics and in-hospital treatment were constructed to evaluate the impact of numbers of risk factors on mortality in critical AMI patients. Results At 2-year follow-up, death was observed in 714 patients (28.7%). A total of 474(19.1%),1541(62%),469(18.9%) patients had 0,1-2,3-4 SMuRFs respectively. No risk group has a significantly higher 2-year mortality (Picture 1). Compared with no risk group, patients with 1-2 and 3-4 SMuRFs were correlated to an HR of 0.72(95%CI, 0.61-0.86) and 0.66(95%CI, 0.52-0.83) for all-cause mortality in the unadjusted model. Interestingly, after multivariate adjustment, less risk factor at AMI onset was still associated with poor prognosis. Conclusions Taking into confounders, critical AMI patients with less risk factors have higher risk in poor prognosis. Further studies on this paradoxical phenomenon are needed.

Background Atherosclerotic plaque hypoxia is detrimental for macrophage function. Prolyl hydroxylases (PHDs) initiate cellular hypoxic responses, possibly influencing macrophage function in plaque hypoxia. Thus, we aimed to elucidate the role of myeloid PHDs in atherosclerosis. Methods Myeloid specific PHD knockout (PHDko) mice were fed high cholesterol diet for 6–12 weeks to induce atherosclerosis. Plaque parameters, e.g. plaque size and macrophage content, were analyzed. Bulk and single cell RNA sequencing was performed on PHD2 BMDMs and plaque macrophages, respectively. Results Aortic root plaque size was augmented 2.6fold in PHD2cko, and 1.4-fold in PHD3ko, but not in PHD1ko mice compared to controls. Macrophage apoptosis was promoted in PHD2cko and PHD3ko mice in vitro and in vivo, via the HIF1α/BNIP3 axis. Bulk and single cell RNA data of PHD2cko bone-marrow-derived macrophages (BMDM) and plaque macrophages, respectively, confirmed these findings and were validated by siRNA silencing. Human plaque BNIP3 mRNA associated with plaque necrotic core, suggesting similar adverse effects. Further, PHD2cko plaques displayed enhanced fibrosis, independent of macrophage MMP activity, collagen secretion or proliferation and of SMC collagen production, or proliferation. Rather, PHD2cko BMDMs enhanced fibroblast collagen secretion in a paracrine manner. Nichenet in silico analysis of macrophage-fibroblast communication predicted SPP1 signaling as regulator, in line with enhanced plaque SPP1 protein content, and SPP1 mRNA in TREM2-foamy plaque macrophages, but not in neutrophils. Conclusion Myeloid PHD2cko and PHD3ko enhanced plaque growth, macrophage apoptosis, and PHD2cko activated paracrine collagen secretion by fibroblasts. Funding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): NWO, Leducq

Early reports indicate that long non-coding RNAs (lncRNAs) are novel regulators of biological responses. However, their role in the human innate immune response, which provides the initial defence against infection, is largely unexplored. To address this issue, here we characterize the long non-coding RNA transcriptome in primary human monocytes using RNA sequencing. We identify 76 enhancer RNAs (eRNAs), 40 canonical lncRNAs, 65 antisense lncRNAs and 35 regions of bidirectional transcription (RBT) that are differentially expressed in response to bacterial lipopolysaccharide (LPS). Crucially, we demonstrate that knockdown of nuclear-localized, NF-κB-regulated, eRNAs (IL1β-eRNA) and RBT (IL1β-RBT46) surrounding the IL1β locus, attenuates LPS-induced messenger RNA transcription and release of the proinflammatory mediators, IL1β and CXCL8. We predict that lncRNAs can be important regulators of the human innate immune response.

Mecp2 is a transcriptional repressor protein that is mutated in Rett syndrome, a neurodevelopmental disorder that is the second most common cause of mental retardation in women. It has been shown that the loss of the Mecp2 protein in Rett syndrome cells alters the transcriptional silencing of coding genes and microRNAs. Herein, we have studied the impact of Mecp2 impairment in a Rett syndrome mouse model on the global transcriptional patterns of long non-coding RNAs (lncRNAs). Using a microarray platform that assesses 41,232 unique lncRNA transcripts, we have identified the aberrant lncRNA transcriptome that is present in the brain of Rett syndrome mice. The study of the most relevant lncRNAs altered in the assay highlighted the upregulation of the AK081227 and AK087060 transcripts in Mecp2-null mice brains. Chromatin immunoprecipitation demonstrated the Mecp2 occupancy in the 5′-end genomic loci of the described lncRNAs and its absence in Rett syndrome mice. Most importantly, we were able to show that the overexpression of AK081227 mediated by the Mecp2 loss was associated with the downregulation of its host coding protein gene, the gamma-aminobutyric acid receptor subunit Rho 2 (Gabrr2). Overall, our findings indicate that the transcriptional dysregulation of lncRNAs upon Mecp2 loss contributes to the neurological phenotype of Rett syndrome and highlights the complex interaction between ncRNAs and coding-RNAs.

Introduction: Long non-coding RNA (lncRNA) have proven to play key roles in cell physiology from nuclear organization and epigenetic remodeling to post-transcriptional regulation. Last decade, gene expression based-classifications have been developed in clear-cell renal cell carcinoma (ccRCC) to identify distinct subtypes of disease and predict patient's outcome. However, there are no current lncRNA comprehensive characterizations in ccRCC. Patients and methodsRNA-sequencing profiles of 475 primary ccRCC samples from the Cancer Genome Atlas (TCGA) were used to assess expressed lncRNA and identify lncRNA-based classification. In addition, integrative analysis was performed to correlate tumor subtypes with copy-number alterations and somatic mutations. ResultsUsing stringent criteria, we identified 1934 expressed lncRNA and assessed their chromatin marks. Unsupervised clustering unravels four lncRNA subclasses in ccRCC associated with distinct clinicopathological and genomic features of this disease. Cluster C2 (23.4%) defines the most aggressive tumours, with the highest Fuhrman grade and stage and the worst overall survival time. Furthermore, cluster C2 is enriched for 9p deletion and chromatin remodeler BAP1 somatic mutations. Interestingly, cluster C4 (7.8%) is related to a tumor subtype arising from the distal tubules of the nephron. Consistent with its distinct ontogeny, cluster C4 is devoid of classical alterations seen in ccRCC, bears frequent 1p deletion and 17q gain, and is enriched for MiTF/TFE translocations. In addition, reexaminations of copy-number data from one side and tumor histology by pathologists from the other side reveal misclassified tumors within C4 cluster including chromophobe RCC and clear cell papillary RCC. ConclusionThis study establishes a foundation for categorizing lncRNA subclasses, which may contribute to understand tumor ontogeny and help predicting patients' outcome in ccRCC.

Introduction Comprehensive cardiac rehabilitation (CR) programs based on the bio-psycho-social approach of the international classification of functioning and disease are carried out to achieve improved prognosis, superior health-related quality of life (HRQL) and social integration. We aimed to identify predictors of returning to work (RTW) and HRQL among cardiovascular risk factors and physical performance as well as patient-reported outcome measures (PROMs) modifiable during CR. Methods We designed a prospective observational multi-center study and enrolled 1,586 patients (2017/18) in 12 German rehabilitation centers regardless of their primary allocation diagnoses (e.g. acute myocardial infarction (AMI), coronary artery bypass grafting (CABG), coronary artery disease (CAD), valvular disease). Besides general data (e.g. age, gender, diagnoses), parameters of risk factor management (e.g. smoking, lipid profile, hypertension, lifestyle change motivation), physical performance (e.g. maximum exercise capacity, endurance training load, 6-min walking distance), and PROMs (e.g. depression, heart-focused anxiety, HRQL, subjective well-being, somatic and mental health, pain, general self-efficacy, pension desire as well as self-assessment of occupational prognosis using several questionnaires) were documented at CR admission and discharge. 6 months after discharge, status of RTW and HRQL (SF-12) were captured by a follow-up (FU) survey and analyzed in multivariable regression models with multiple imputation of missing values. Results Out of the study participants, 1,262 patients (54±7 years, 77% men) responded to the follow-up survey and could be analyzed regarding the outcome parameters. Most of them were assigned to CR primarily due to AMI (40%) or CAD without myocardial infarction (18%), followed by heart valve diseases in 12% of patients and CABG (8%). 864 patients (69%) returned to work within the follow-up period. Pension desire, negative self-assessed occupational prognosis, heart-focussed anxiety, major life events, smoking and heart failure were negatively associated with RTW, while higher endurance training load, HRQL and work stress were positively associated (Figure 1). HRQL after 6 months was determined more by PROMs (e.g. pension desire, heart-focused anxiety, physical/mental HRQL in SF-12, physical/mental health in indicators of rehab-status questionnaire (IRES-24), stress, well-being in the World Health Organization well-being index and self-efficacy expectations) than by clinical parameters or physical performance. Conclusions Patient-reported outcome measures predominantly influenced RTW and HRQL in heart-disease patients, whereas patients' pension desire and heart-focussed anxiety had a dominant impact on all investigated endpoints. Therefore, the multi-component CR approach focussing on psychosocial support is crucial for subjective health prognosis and occupational resumption. Figure 1. Predictors of returning to work Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): German pension insurance

RNA polyadenylation is crucial for RNA maturation, stability and function, with poly(A) tail lengths significantly influencing mRNA translation, efficiency and decay. Here, we provide a step-by-step protocol to perform Nanopore 3' end-capture sequencing (Nano3P-seq), a nanopore-based cDNA sequencing method to simultaneously capture RNA abundances and tail-composition and tail-length estimates at single-molecule resolution. Taking advantage of a template switching-based protocol, Nano3P-seq can sequence any RNA-derived molecule from its 3' end, regardless of its polyadenylation status, without the need for PCR amplification or RNA adapter ligation. We provide an updated Nano3P-seq protocol that is compatible with R10.4 flow cells, as well as compatible software for poly(A) tail length and content prediction, which we term 'PolyTailor'. We demonstrate that PolyTailor provides accurate estimates of transcript abundances and tail lengths and composition, while capturing both coding and noncoding RNA biotypes, including mRNAs, small nucleolar RNAs and ribosomal RNAs. Nano3P-seq can be applied to RNA samples prepared by using different methods (e.g., poly(A)-selected, ribodepleted or total RNA) and can be completed in 1 day. The protocol requires experience in molecular biology techniques and nanopore sequencing library preparation and basic knowledge of Linux Bash syntax and R programming. This protocol makes Nano3P-seq accessible and easy to implement by future users aiming to study the tail dynamics and heterogeneity of both coding and noncoding transcriptomes in a comprehensive and reproducible manner.

Introduction: Serrated colon polyps are found in 15-20% of patients undergoing screening colonoscopy and may account for up to a third of all colorectal cancers (CRC). Sessile serrated adenomas/polyps (SSA/Ps) have a high risk for development into CRC. However, the molecular biology of serrated pathway to cancer is poorly understood. We describe multiple unique non-coding RNA differentially expressed in SSA/Ps and provide an insight into the potential pathways to serrated carcinoma. Methods: Small RNA sequencing was performed on 66 colon RNA samples including 13 SSA/P's, 9 Hyperplastic polyps, 7 Adenomas, 22 uninvolved colon and 15 control colon. The small (< 200 bases) RNA transcriptome of SSA/Ps and other polyp types were characterized to identify unique non-coding RNA expression in SSA/Ps. Gene targets for each of our differentially expressed microRNAs (miRNAs) were identified using multiple miRNA databases. Results: We found 104 small RNAs including 53 miRNA, 46 small nucleolar RNA and 5 nuclear RNA differentially expressed (Fold change >1.5, False discovery rate MIR31) was most differentially expressed (>6 fold) followed by MIR135B expression (>4-fold) in SSA/Ps. MIR378A was downregulated by twofold in SSA/Ps. We identified 14 gene targets for MIR378A that were statistically overexpressed in miRNA sequencing data, four of which are involved in insulin growth factor signaling. Conclusion: We describe a comprehensive analysis of the small non-coding RNA transcriptome in SSA/Ps and HPs. Increased expression of MIR31 has beendescribed in SSA/Ps and our data is consistent with recent studies. MIR135B overexpression and its role in the APC/Wnt pathway has been described in CRC and our study suggests disruption of Wnt signaling possibly playing a role in the serrated pathway. MIR378A is a tumor suppressor gene and we present the first evidence of decreased expression of MIR378A in SSA/Ps. 4 of the 14 MIR378A gene targets identified in our study are important in insulin growth factor signaling pathway which may be contributing to serrated cancer development. Our data provides insights into small RNA expression unique to SSA/Ps and potential oncological pathways which may help understand the molecular biology of progression of SSA/Ps into CRC.

BackgroundGlobally, most deaths result from cardiovascular diseases, particularly ischemic heart disease. COVID-19 affects the heart, worsening existing heart conditions and causing myocardial injury. The mechanistic link between COVID-19 and acute myocardial infarction (AMI) is still being investigated to elucidate the underlying molecular perspectives.MethodsGenetic risk assessment was conducted using two-sample Mendelian randomization (TSMR) to determine the causality between COVID-19 and AMI. Weighted gene co-expression network analysis (WGCNA) and machine learning were used to discover and validate shared hub genes for the two diseases using bulk RNA sequencing (RNA-seq) datasets. Additionally, gene set enrichment analysis (GSEA) and single-cell RNA-seq (scRNA-seq) analyses were performed to characterize immune cell infiltration, communication, and immune correlation of the hub genes. To validate the findings, the expression patterns of hub genes were confirmed in clinical blood samples collected from COVID-19 patients with AMI.ResultsTSMR did not find evidence supporting a causal association between COVID-19 or severe COVID-19 and AMI. In the bulk RNA-seq discovery cohorts for both COVID-19 and AMI, WGCNA’s intersection analysis and machine learning identified TLR4 and ABCA1 as significant hub genes, demonstrating high diagnostic and predictive value in the RNA-seq validation cohort. Single-gene GSEA and single-sample GSEA (ssGSEA) revealed immune and inflammatory roles for TLR4 and ABCA1, linked to various immune cell infiltrations. Furthermore, scRNA-seq analysis unveiled significant immune dysregulation in COVID-19 patients, characterized by altered immune cell proportions, phenotypic shifts, enhanced cell-cell communication, and elevated TLR4 and ABCA1 in CD16 monocytes. Lastly, the increased expression of TLR4, but not ABCA1, was validated in clinical blood samples from COVID-19 patients with AMI.ConclusionNo genetic causal link between COVID-19 and AMI and dysregulated TLR4 and ABCA1 may be responsible for the development of immune and inflammatory responses in COVID-19 patients with AMI.

Background Rapid reduction of heart rate (HR) is the most evident physiological response of vagal nerve stimulation (VNS). Since HR reduction is the most potent factor to decrease myocardial oxygen consumption rate (MVO2), the appropriate VNS can exert cardio-protective effects. It is also known that VNS reduces inflammation, oxidative stress, and sympathetic overload. In addition, the VNS during ischemia-reperfusion is known to attenuate myocardial damage by studies in various animal species. Despite the presence of preclinical evidence of VNS benefits, the lack of the device has limited the translation of this technology to clinical practice. We have recently developed an intravenous VNS catheter (JOHAKU, Neuroceuticals Inc.) that can stimulate the right vagal nerve via superior vena cava (SVC) (Figure 1) on temporary basis. Purpose We aimed to confirm the feasibility of JOHAKU as a device to modulate heart rate and MVO2 rapidly by a canine experiment. Methods In eight beagle dogs, JOHAKU was inserted from the right femoral vein and placed at the SVC level. The stimulation intensity was adjusted to 10–20 V (20 Hz). We simultaneously recorded electrocardiogram and intraarterial blood pressure (BP). In three of eight dogs, we measured the left anterior descending coronary artery flow and oxygen saturations of arterial and coronary sinus blood to calculate MVO2. We compared HR, BP, and MVO2 during JOHAKU stimulation to ones at baseline. Results As shown in Figure 2, JOHAKU attenuated HR immediately after stimulation. Compared with baseline, JOHAKU significantly reduced HR (baseline: 135±13 vs. 5 min on stimulation: 107±13 bpm, p&amp;lt;0.05), and did not affect mean BP significantly (96.2±22.8 vs. 89.4±26.6 mmHg, P=0.59). HR promptly recovered to baseline level after JOHAKU stopped. JOHAKU also reduced MVO2 (0.57±0.43 vs. 0.48±0.38 ml/min, p&amp;lt;0.05). Conclusion JOHAKU rapidly attenuated cardiac metabolism burden via the rapid HR reduction. The controllability of HR by JOHAKU without affecting BP enables us to apply the VNS even for patients with hemodynamic instability, such as heart failure and acute myocardial infarction. Funding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Japan Agency for Medical and Research Development

Introduction and aim Adverse ventricular remodelling and heart failure (HF) are frequent complications of acute myocardial infarction (MI), but no reliable methods are currently available for prediction. MicroRNAs (miRNAs) can be used as biomarkers and they play a crucial role in multiple diseases onset and evolution. The aim of our study was to identify and validate a panel of serum miRNAs with differential expression in MI and test their prognostic ability to predict chronic HF. Methods Sequential approach with several models: 1) Initial selection of miRNAs by a 762 miRNA hybridization array with samples from 12 patients with acute MI and 10 age and sex-matched controls; 2) Validation of selected miRNAs in a prospective study with consecutive enrolment of patients with type 1 MI at the Cardiac ICU of a tertiary hospital; two independent cohorts were included (2013–2014 and 2017–2019) with clinical and CV imaging assessment at baseline and long-term follow-up; serum miRNAs and other biomarkers (BNP, NT-proBNP, troponin, etc) were studied; the composite HF-related outcome included HF hospitalizations and CV mortality; 30 age and sex-adjusted controls were studied for miRNAs comparison; miRNA signature study was completed with 3) in vitro model of hypoxia ±nutrient supply deprivation in H9C2 cardiomyocytes, 4) in vivo rat model of ischemia-reperfusion and 5) permanent coronary occlusion. All procedures were approved by the Ethics Committee (refs. 175/13 &amp; 061/16), the Animals Ethics Committee (ref. 298/16) and complied with the Declaration of Helsinki. All patients provided written informed consent. Results 14 miRNAs were initially identified with highest differentiation between MI and controls. In the validation study, 311 patients with MI were enrolled, mean age 60±15, 81% male; ST elevation MI 65%, GRACE risk score 144±45, Killip class ≥II 23%, LVEF&amp;lt;50% at discharge 46% and NT-proBNP 3091±5997 pg/ml. miR-210, miR-21, miR-23 and miR-221 discriminated between patients with more severe form of MI presentation by Killip class (figure 1-B); in addition, GRACE risk score statistically correlated with miR-21, miR-221, miR-27, miR-93, miR-23, miR-148, miR-107, miR-210 and miR-144. Among markers of acute HF, miR-21, miR-23, miR-27, miR-210 and miR-221 significantly correlated with NT-proBNP (figure 1-C) and BNP levels. In parallel, miR-210, miR-21, miR-23, miR-106, miR-221, miR-27 and miR-93 correlated with left ventricular ejection fraction (LVEF). Finally, miR-21, miR-23, miR-210 and miR-221 associated with patients' long-term survival free of long-term HF-related events. Both in vitro and in vivo experimental models served as confirmation of the importance of these outlined miRNAs in cardiomyocyte response to ischemic stress and ventricular remodelling. Conclusions We identified a signature of microRNAs associated with adverse ventricular remodelling and poor long-term outcome after MI, that could serve as biomarkers involved in HF progression. Figure 1 Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): Instituto de Salud Carlos III

Background Obstructive sleep apnea (OSA) is closely related to the incidence and progression of coronary artery disease (CAD), but the mechanisms linking OSA and CAD are unclear. C1q/TNF-related protein-9 (CTRP9) is a novel adipokine that protects the heart against ischemic injury and ameliorates cardiac remodeling. Purpose We aimed to ascertain the clinical relevance of CTRP9 with OSA prevalence in patients with CAD. Methods From August 2016 to March 2019, consecutive eligible patients with CAD (n=154; angina pectoris, n=88; acute myocardial infarction [AMI], n=66) underwent cardiorespiratory polygraphy during hospitalization. OSA was defined as an apnea-hypopnea index (AHI) ≥15 events h–1. Plasma CTRP9 concentrations were measured by ELISA method. Results OSA was present in 89 patients (57.8%). CTRP9 levels were significantly decreased in the OSA group than in the non-OSA group (4.7 [4.1–5.2] ng/mL vs. 4.9 [4.4–6.0] ng/mL, P=0.003). The difference between groups was only observed in patients with AMI (3.0 [2.3–4.9] vs. 4.5 [3.2–7.9], P=0.009), but not in patients with AP (5.0 [4.7–5.3] ng/mL vs. 5.1 [4.7–5.9] ng/mL, P=0.571) (Figure 1). Correlation analysis showed that CTRP9 levels were negatively correlated with AHI (r=−0.238, P=0.003) and oxygen desaturation index (r=−0.234, P=0.004), and positively correlated with left ventricular ejection fraction (r=0.251, P=0.004) in all subjects. Multivariate analysis showed that male gender (OR 3.099, 95% CI 1.029–9.330, P=0.044), body mass index (OR 1.148, 95% CI 1.040–1.268, P=0.006), and CTRP9 levels (OR 0.726, 95% CI 0.592–0.890, P=0.002) were independently associated with the prevalence of OSA. Conclusions Plasma CTRP9 levels were independently related to the prevalence of OSA in patients with CAD, suggesting that CTRP9 might play a role in the pathogenesis of CAD exacerbated by OSA. Figure 1. CTRP9 levels in OSA and non-OAS groups Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): National Natural Science Foundation of China

Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): grant of the President of the Russian Federation Background. In recent years a group of patients with acute myocardial infarction without obstructive coronary artery disease (MINOCA) is of interest. In this group of patients, there is a lack of information related to myocardial perfusion, blood flow and reserve, which could be important in term of risk and prognosis assessment. Purpose. To assess the global and regional myocardial blood flow and coronary flow reserve impairment in MINOCA patients compared to cardiac magnetic resonance (CMR). Methods. The study group comprised 28 patients (18 male, mean 62.9 ± 10.7years) with acute coronary syndrome. Based on ICA results all patients were divided into two groups: 1) MINOCA (n = 11); 2) patients with myocardial infarction with obstructive CAD (MICAD) (n = 17). A total of 12/17 (71%) MICAD patients underwent PCI with stenting; in 2/12 cases two arteries were revascularized. Within 7-10 days after admission to the hospital all patients underwent SPECT myocardial perfusion scintigraphy (MPS) with the assessment of standard indices (SSS, SRS, SDS) and quantitative parameters: rest MBF (rMBF), stress MBF (sMBF) and CFR (on CZT gamma-camera). Also, all patients underwent CMR with evaluation of infarct size (IF), microvascular obstruction (MVO), size of myocardial edema (ME) and transmural extent of myocardial infarction (TE). Results Visual analysis showed that SSS differed significantly (p = 0.001) among MINOCA and MICAD groups: 5 (3;6) vs 10 (5;13), respectively. According to the quantitative MPS data analysis, global sMBF and CFR values were significantly higher in MINOCA than in MICAD patients group: 1.2 (0.82;1.69) ml/min/g vs 0.62 (0.52;0.9) ml/min/g; 1.96 (1.23;2.42) vs 1.16 (0.98;1.64) , respectively. CMR indices such as IS, ME and TE were significantly lower in MINOCA compared to MICAD patients: 1.6 (0;7.8) vs 14.8 (6.3;22.6)%; 8(0;14) vs 18.5 (10;23)%; 0 (0;5) vs 18 (10;25), respectively. Seven(40%) MICAD group patients showed MVO, whereas there were no such patients in MINIOCA group. The regional analysis revealed that sMBF and CFR were significantly lower in LV regions characterized by myocardial injury compared to non-injured regions (based on CMR data): 0.78 (0.51;0.94) ml/min/g vs 1.11 (0.82;1.4) and 1.22 (0.91;1.74) vs 1.52 (1.14;2.42), respectively. Also, we found out significant correlation between regional quantitative SPECT indices and transmural extent of myocardial infarction evaluated by CMR: r=–0.4 for sMBF; r=–0.35 for CFR (p &amp;lt; 0.05). Conclusion. The analysis of quantitative MPS SPECT parameters showed that such approach allows identifying MBF and CFR disturbances both on global and regional level. These results showed that MINOCA patients characterized by mild reduction of myocardial blood flow and perfusion assessed visually and quantitatively. It means that despite the absence of obstructive coronary artery lesion this group of patients has more pronounced risk of cardiac events and need more aggressive observation and treatment.

Background Cell therapy has emerged as a promising additional therapy after myocardial infarction (MI). We have previously shown that interactions between risk factors and cell therapy influence efficacy, suggesting that only certain patient populations benefit from cell therapy after MI. Purpose We aim to generate and validate a prediction model for cell therapy after MI for both functional outcome and clinical endpoints. Methods Data from the REPAIR-AMI and REGENERATE-AMI trial were used to model the effect of autologous bone marrow-derived mononuclear cell transplantation after acute MI. First, a prediction model was generated based on the REPAIR-AMI trial. The outcome measure for the model was difference in ejection fraction (EF) over time. Prespecified variables of interest were age, sex, weight, hypertension, hyperlipidaemia, diabetes, history of smoking and baseline EF, taking into account their interaction per therapy (placebo vs cells). This model resembles the gain in function over time after cell therapy, compared to the placebo group. Validation of this model was performed in the REGENERATE-AMI trial dataset, to test whether interactions that influence the therapy are similar across trials. Finally, the model was tested for its ability to predict clinical outcome (i.e. MACE) in REPAIR-AMI. Results The generated prediction model on REPAIR-AMI data included multiple interactions with therapy. Internally, the model functioned appropriately to predict the difference in EF (n=186, R2=0.17, p=9x10–9). In the validation step, our model showed similar predictive capacity in REGENERATE-AMI (n=94, R2=0.19, p=1.4x10–5) specifically in the cell therapy group (R2=0.31, p=1.9x10–5) (see Figure). A multivariable analysis on the combined datasets revealed that patients with a smoking history or low baseline EF were more likely to have an increased functional response after cell therapy. Applying this prediction model to clinical endpoint data showed that a generated benefit score (based on cardiac function) translates to differences in event rate. The high benefit group yielded a significantly different event rate between groups (45% in placebo vs 15% in cell therapy group, p=0.001). Conclusion Based on real life clinical data of two autologous cell therapy trials we have generated a clinically useful prediction model for the response to cell therapy after MI. Particular factors (smoking, baseline EF) showed relevant interactions. Not only functional outcome, but also clinical endpoints are adequately predicted by a model-derived benefit score, indicating that specific patient groups might benefit more from cell therapy. Ideally, this will be validated prospectively in large trials, like the BAMI trial (NCT01569178) and ultimately implemented in future clinical trial design to identify these patient groups upfront. A future analysis of the BAMI trial data will follow after its primary publication, using our generated prediction model. Figure 1. Validation in REGENERATE-AMI Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): CVON HUSTCARE, Alexandre Suerman stipend UMC Utrecht