Translational and Mechanistic Study about Beta-1-Adrenergic Receptor Modulation on Neutrophils as a Therapy against Ischemia/Reperfusion Injury

Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Bioquímica. Fecha de Lectura: 24-02-2023This work received funding from the Instituto de Salud Carlos III (ISCIII; PI16/02110 and PT20/00044), the European Regional Development Fund (ERDF) “A way of making Europe", the Comunidad de Madrid (S2017/BMD-3867 RENIM-CM) cofunded with European structural and investment funds and by Agencia Estatal de Investigación (PID2019‐110369RB‐I00). The CNIC is supported by the ISCIII, the Ministerio de Ciencia e Innovación and the Pro CNIC Foundation and is a Severo Ochoa Center of Excellence (CEX2020-001041-S

Papel de la toxina pertussis y adenilato ciclasa en la prevalencia intracelular de bordetella pertussis y desarrollo de nuevas vacunas

La tos ferina es una enfermedad infecciosa originada por Bordetella pertussis, que constituye uno de los principales problemas de salud pública mundial en la actualidad. En países desarrollados, a pesar de la alta cobertura de vacunación en la infancia, se ha reportado un aumento de casos en ciertos grupos de edad, como adolescentes y adultos jóvenes, así como lactantes pequeños, presentando éstos las mayores tasas de hospitalización, complicaciones graves y mortalidad. Sin embargo, poco se conoce acerca de las formas y mecanismos de persistencia de este patógeno. En los últimos años, sucesivos estudios han postulado la existencia de una fase intracelular en la que dos de las principales toxinas de Bordetella pertussis, como la toxina pertussis (PT) y adenilato ciclasa (TAC), parecen desempeñar un papel fundamental, contribuyendo así a la prevalencia bacteriana. Todo ello, junto a las limitaciones de las actuales estrategias de vacunación basadas en vacunas de carácter acelular, supone la necesidad de proseguir la investigación en búsqueda de vacunas más apropiadas para neonatos y con mejor inmunogenicidad

Metoprolol exerts a non-class effect against ischaemia-reperfusion injury by abrogating exacerbated inflammation.

Clinical guidelines recommend early intravenous β-blockers during ongoing myocardial infarction; however, it is unknown whether all β-blockers exert a similar cardioprotective effect. We experimentally compared three clinically approved intravenous β-blockers. Mice undergoing 45 min/24 h ischaemia-reperfusion (I/R) received vehicle, metoprolol, atenolol, or propranolol at min 35. The effect on neutrophil infiltration was tested in three models of exacerbated inflammation. Neutrophil migration was evaluated in vitro and in vivo by intravital microscopy. The effect of β-blockers on the conformation of the β1 adrenergic receptor was studied in silico. Of the tested β-blockers, only metoprolol ameliorated I/R injury [infarct size (IS) = 18.0% ± 0.03% for metoprolol vs. 35.9% ± 0.03% for vehicle; P < 0.01]. Atenolol and propranolol had no effect on IS. In the three exacerbated inflammation models, neutrophil infiltration was significantly attenuated only in the presence of metoprolol (60%, 50%, and 70% reductions vs. vehicle in myocardial I/R injury, thioglycolate-induced peritonitis, and lipopolysaccharide-induced acute lung injury, respectively). Migration studies confirmed the particular ability of metoprolol to disrupt neutrophil dynamics. In silico analysis indicated different intracellular β1 adrenergic receptor conformational changes when bound to metoprolol than to the other two β-blockers. Metoprolol exerts a disruptive action on neutrophil dynamics during exacerbated inflammation, resulting in an infarct-limiting effect not observed with atenolol or propranolol. The differential effect of β-blockers may be related to distinct conformational changes in the β1 adrenergic receptor upon metoprolol binding. If these data are confirmed in a clinical trial, metoprolol should become the intravenous β-blocker of choice for patients with ongoing infarction.Ministry of Science and Innovation (‘RETOS 2019’ grant N_ PID2019-107332RB-I00), Instituto de Salud Carlos III (ISCIII; PI16/02110), and European Regional Development Fund (# AC16/00021), Comunidad de Madrid (S2017/BMD-3867 RENIM-CM). B.I. is supported by an ERCCoG grant (819775). E.O. is supported by funds from the Comunidad de Madrid Programa de Atraccion de Talento (2017-T1/BMD-5185). A.C-M. and R.V-G are supported by fellowships from the Ministerio de Ciencia e Innovacion (MCN) and ISCIII (FPU2017/01932 and PFIS FI17/00045). D.V.L. is supported by an Iniciativa de Empleo Juvenil grant (PEJ-2017-TL/BMD-6463) from the Comunidad de Madrid. The CNIC is supported by the ISCIII, the MCN, and the Pro CNIC Foundation and is a Severo Ochoa Center of Excellence (SEV-2015-0505).S

Metoprolol in Critically Ill Patients With COVID-19.

Severe coronavirus disease-2019 (COVID-19) can progress to an acute respiratory distress syndrome (ARDS), which involves alveolar infiltration by activated neutrophils. The beta-blocker metoprolol has been shown to ameliorate exacerbated inflammation in the myocardial infarction setting. The purpose of this study was to evaluate the effects of metoprolol on alveolar inflammation and on respiratory function in patients with COVID-19-associated ARDS. A total of 20 COVID-19 patients with ARDS on invasive mechanical ventilation were randomized to metoprolol (15 mg daily for 3 days) or control (no treatment). All patients underwent bronchoalveolar lavage (BAL) before and after metoprolol/control. The safety of metoprolol administration was evaluated by invasive hemodynamic and electrocardiogram monitoring and echocardiography. Metoprolol administration was without side effects. At baseline, neutrophil content in BAL did not differ between groups. Conversely, patients randomized to metoprolol had significantly fewer neutrophils in BAL on day 4 (median: 14.3 neutrophils/µl [Q1, Q3: 4.63, 265 neutrophils/µl] vs median: 397 neutrophils/µl [Q1, Q3: 222, 1,346 neutrophils/µl] in the metoprolol and control groups, respectively; P = 0.016). Metoprolol also reduced neutrophil extracellular traps content and other markers of lung inflammation. Oxygenation (PaO2:FiO2) significantly improved after 3 days of metoprolol treatment (median: 130 [Q1, Q3: 110, 162] vs median: 267 [Q1, Q3: 199, 298] at baseline and day 4, respectively; P = 0.003), whereas it remained unchanged in control subjects. Metoprolol-treated patients spent fewer days on invasive mechanical ventilation than those in the control group (15.5 ± 7.6 vs 21.9 ± 12.6 days; P = 0.17). In this pilot trial, intravenous metoprolol administration to patients with COVID-19-associated ARDS was safe, reduced exacerbated lung inflammation, and improved oxygenation. Repurposing metoprolol for COVID-19-associated ARDS appears to be a safe and inexpensive strategy that can alleviate the burden of the COVID-19 pandemic.Mr Clemente-Moragón is supported by a fellowship from the Ministerio de Ciencia e Innovación (FPU2017/01932). The CNIC is supported by the ISCIII, the Ministerio de Ciencia e Innovación, and the Pro CNIC Foundation. Dr Ibáñez is supported by the European Commission (ERC-CoG grant No 819775) and by the Spanish Ministry of Science and Innovation (MCN; “RETOS 2019” grant No PID2019- 107332RB-I00). Dr Oliver is supported by funds from the Comunidad de Madrid Programa de Atracción de Talento (2017-T1/BMD-5185). All other authors have reported that they have no relationships relevant to the contents of this paper to disclose.S

Translational and Mechanistic Study about Beta-1-Adrenergic Receptor Modulation on Neutrophils as a Therapy against Ischemia/Reperfusion Injury

[EN] Acute myocardial infarction and ischemic stroke are the two most frequent life-threatening presentations of atherosclerosis. These two conditions are the leading causes of morbidity and mortality worldwide. In both cases, the main determinant of poor outcome is the extent of irreversible injury. The mainstay treatment of acute myocardial infarction and ischemic stroke is timely reperfusion of the occluded artery to restore blood flow. However, despite being essential for tissue salvage, evidence shows that reperfusion itself triggers a set of potentially deleterious events that paradoxically contribute to final infarct size. From the different mechanisms involved in reperfusion-related injury, microvascular obstruction and acute inflammatory response play a central role. The beta-1-selective blocker metoprolol has been demonstrated to reduce myocardial infarct size through its inhibitory effect on neutrophils. Based on the cardioprotective effect of metoprolol injection, among other effects (e.g. antiarrhythmic effect); current clinical practice guidelines recommend early intravenous administration of beta-blockers to patients with an ongoing acute myocardial infarction. While different beta-blockers exist (with clear pharmacological differences among them), given the lack of comparative studies, clinical practice guidelines do not recommend on over another. While the adjuvant pharmacological treatment to reperfusion is well developed for acute myocardial infarction, treatment of stroke is currently restricted to reperfusion without coadjuvant therapies. Within this thesis, the biological effects of the different clinically available intravenous beta-blockers on different acute conditions have been tested. This work presents evidence that metoprolol has a particular action on neutrophils during exacerbated inflammation (acute myocardial infarction, peritonitis and acute lung injury) that affords a cardioprotection not provided by other beta-blockers, such as atenolol or propranolol. Besides, herein it is shown how these effects attributed to metoprolol are translated into protection in other different and complex pathological scenarios, in which neutrophils are shown to play a key role, such as ischemic stroke or acute respiratory distress syndrome. Although larger clinical studies are needed to confirm these results, the identification of metoprolol’s mechanism of action through neutrophil stunning has broadened the potential of an old drug as a therapeutic strategy against prevalent diseases for which adjuvant strategies are thoroughly limited (e.g. ischemic stroke and acute respiratory distress syndrome). Likewise, this work contributes to highlight the relevance of drug repurposing, as a highly efficient, time saving, low-cost and minimum risk of failure strategy, so as to speed up the slow development of a molecule from bench to market. Noteworthy, this thesis includes the execution of a clinical study in patients with severe COVID-19 (a condition characterized by acute respiratory distress syndrome with massive lung inflammatory response). The contribution of basic and translational research to the fight against this terrible pandemic is a role model of how science is vital in our society.[ES] El infarto agudo de miocardio y el ictus isquémico son dos de las presentaciones de mayor frecuencia y potencialmente fatales de aterosclerosis. Estas dos condiciones son consideradas dos de las mayores causas de morbimortalidad a nivel mundial. En ambos casos, el principal determinante de un mal pronóstico es la extensión de la lesión irreversible. El principal tratamiento del infarto agudo de miocardio y el accidente cerebrovascular isquémico es la reperfusión de la arteria ocluida con el fin de restablecer el flujo sanguíneo. A pesar de ser esencial para la recuperación del tejido, la evidencia muestra que la reperfusión en sí desencadena una serie de eventos potencialmente nocivos que, paradójicamente, contribuyen al tamaño final del infarto. De los diferentes mecanismos implicados en la lesión por reperfusión, la obstrucción microvascular y la respuesta inflamatoria aguda juegan un papel central. La administración intravenosa previa a la reperfusión del antagonista beta-1-adrenérgico metoprolol ha demostrado ser capaz de reducir el tamaño del infarto de miocardio a través de su efecto inhibidor sobre los neutrófilos. Basándose en parte en el efecto cardioprotector de la inyección intravenosa de metoprolol, entre otros efectos (por ejemplo, su efecto antiarrítmico), las guías de práctica clínica actuales recomiendan la administración intravenosa temprana de beta-bloqueantes (como clase farmacoterapéutica) en pacientes con un infarto agudo de miocardio. Si bien existen diferentes beta-bloqueantes (con claras diferencias farmacológicas entre ellos), dada la falta de estudios comparativos, las guías de práctica clínica no recomiendan uno sobre otro. El tratamiento farmacológico adyuvante a la reperfusión está bien desarrollado para el infarto agudo de miocardio; sin embargo, el tratamiento actual del ictus isquémico se restringe a la reperfusión sin terapias coadyuvantes. En la presente tesis doctoral se han probado los efectos biológicos de los diferentes beta-bloqueantes intravenosos clínicamente disponibles en diferentes condiciones agudas. Este trabajo presenta evidencias de que el metoprolol intravenoso ejerce una acción única sobre los neutrófilos durante la inflamación exacerbada (infarto agudo de miocardio, peritonitis y lesión pulmonar aguda) que proporciona una cardioprotección no asociada al uso de otros beta-bloqueantes, como el atenolol o el propranolol. Además, en la presente tesis se muestra cómo estos efectos atribuidos al metoprolol se traducen en protección en otros escenarios patológicos diferentes y complejos, en los que los neutrófilos muestran un papel clave, como el ictus isquémico o el síndrome de distrés respiratorio agudo. Aunque se necesitan estudios clínicos más amplios para confirmar estos resultados, la identificación del mecanismo de acción de metoprolol a través del aturdimiento de neutrófilos ha ampliado el potencial de un fármaco antiguo como estrategia terapéutica frente a enfermedades prevalentes para las que las estrategias adyuvantes son limitadas (por ejemplo, accidente cerebrovascular isquémico y enfermedades respiratorias agudas). Asimismo, este trabajo contribuye a resaltar la relevancia del reposicionamiento de fármacos, como una estrategia de alta eficiencia, ahorro de tiempo, bajo coste y mínimo riesgo de fracaso, con el objetivo de acelerar el lento desarrollo de una molécula desde el laboratorio hasta el mercado. Por último, cabe destacar que esta tesis doctoral incluye la ejecución de un estudio clínico en pacientes con COVID-19 grave (una condición caracterizada por síndrome de dificultad respiratoria aguda con respuesta inflamatoria pulmonar masiva). La contribución de la investigación básica y traslacional a la lucha contra esta terrible pandemia es un modelo a seguir de cómo la ciencia es vital en nuestra sociedad.Instituto de Salud Carlos III (ISCIII; PI16/02110 and PT20/00044), the European Regional Development Fund (ERDF) “A way of making Europe", the Comunidad de Madrid (S2017/BMD-3867 RENIM-CM) cofunded with European structural and investment funds and by Agencia Estatal de Investigación (PID2019‐110369RB‐I00). The CNIC is supported by the ISCIII, the Ministerio de Ciencia e Innovación and the Pro CNIC Foundation and is a Severo Ochoa Center of Excellence (CEX2020-001041-S). Agustín Clemente Moragón is the beneficiary of an FPU fellowship from the Ministerio de Ciencia e Innovación (FPU2017/01932). In the course of his Doctoral Thesis, Agustín Clemente Moragón did a 6-months research stay (October 2021 to March 2022) at Dr. Filip Swirski’s Lab in the Cardiovascular Research Institute at Icahn School of Medicine at Mount Sinai in New York (US), with funding from the Ministerio de Ciencia e Innovación (EST21/00633) and The Company of Biologists (DMMTF2110609).Peer reviewe

Anthracycline Cardiotoxicity Induces Progressive Changes in Myocardial Metabolism and Mitochondrial Quality Control

Dr Ibáñez is supported by the European Commission (grants ERC-CoG 819775 and H2020-HEALTH 945118), the Spanish Ministry of Science, Innovation and Universities (grant PID2022-140176OB-I00), and Comunidad de Madrid through the Red Madrileña de Nanomedicina en Imagen Molecular (grant P2022/BMD-7403 RENIM-CM). Dr DíazGuerra’s PhD fellowship is funded by the Spanish Association Against Cancer. Dr Oliver is a Ramón y Cajal fellow (grant RYC2020-028884-I) funded by MCIN/AEI/10.13039/501100011033 and by “ESF Investing in Your Future.” Centro Nacional de Investigaciones Cardiovasculares is supported by Instituto de Salud Carlos III, Ministerio de Ciencia e Innovación, and the Pro CNIC Foundation and is a Severo Ochoa Center of Excellence (grant CEX2020-001041-S funded by MICIN/AEI/ 10.13039/501100011033). The authors have reported that they have no relationships relevant to the contents of this paper to disclose.S

Oxidative Post-translational Protein Modifications upon Ischemia/Reperfusion Injury.

While reperfusion, or restoration of coronary blood flow in acute myocardial infarction, is a requisite for myocardial salvage, it can paradoxically induce a specific damage known as ischemia/reperfusion (I/R) injury. Our understanding of the precise pathophysiological molecular alterations leading to I/R remains limited. In this study, we conducted a comprehensive and unbiased time-course analysis of post-translational modifications (PTMs) in the post-reperfused myocardium of two different animal models (pig and mouse) and evaluated the effect of two different cardioprotective therapies (ischemic preconditioning and neutrophil depletion). In pigs, a first wave of irreversible oxidative damage was observed at the earliest reperfusion time (20 min), impacting proteins essential for cardiac contraction. A second wave, characterized by irreversible oxidation on different residues and reversible Cys oxidation, occurred at late stages (6-12 h), affecting mitochondrial, sarcomere, and inflammation-related proteins. Ischemic preconditioning mitigated the I/R damage caused by the late oxidative wave. In the mouse model, the two-phase pattern of oxidative damage was replicated, and neutrophil depletion mitigated the late wave of I/R-related damage by preventing both Cys reversible oxidation and irreversible oxidation. Altogether, these data identify protein PTMs occurring late after reperfusion as an actionable therapeutic target to reduce the impact of I/R injury.This study was supported by competitive grants from the MCIN/AEI/10.13039/501100011033/ and FEDER Una manera de hacer Europa (PID2021-122348NB-I00 and PID2022-140176OB-I00), the MCIN/AEI/10.13039/501100011033 the Unión Europea NextGenerationEU/PRTR (PLEC2022- 009235, and PLEC2022-009298), the Comunidad de Madrid (Programa S2022/BDM-7333-CM, INMUNOVAR, P2022/BMD-7333, and RENIM-CM, P2022/BMD-7403), “la Caixa” Banking Foundation (HR17- 00247, LCF/PR/HP22/52320018, and LCF/PR/HR22/52420019), and the European Commission (ERC Consolidator Grant “MATRIX”, 819775). Rodrigo Fernandez-Jimenez is the recipient of grant PI22/01560 funded by the ISCIII-Fondo de Investigación Sanitaria and co-funded by the European Union. Aleksandra Binek and Navratan Bagwan were FP7-PEOPLE-2013-ITN-Cardionext fellows from the European Union’s Seventh Framework Programme (Marie Curie Actions). Celia Castans holds a Formación del Profesorado Universitario (FPU14/05292) fellowship from the Spanish Ministry of Education, Culture and Sports. Eduardo Oliver holds a Ramón y Cajal fellowship (RYC2020-028884- I) and granted by the MCIN/AEI/10.13039/501100011033/ and FEDER Una manera de hacer Europa (PID2021-133167OB-100) and by “ESF Investing in your future”. The CNIC is supported by the Instituto de Salud Carlos III (ISCIII), the Ministerio de Ciencia e Innovación (MCIN), and the Pro CNIC Foundation and is a Severo Ochoa Center of Excellence (grant CEX2020-001041-S funded by MICIN/AEI/10.13039/501100011033).S

Beta-3 adrenergic receptor overexpression reverses aortic stenosis-induced heart failure and restores balanced mitochondrial dynamics

Aortic stenosis (AS) is associated with left ventricular (LV) hypertrophy and heart failure (HF). There is a lack of therapies able to prevent/revert AS-induced HF. Beta3 adrenergic receptor (β3AR) signaling is beneficial in several forms of HF. Here, we studied the potential beneficial effect of β3AR overexpression on AS-induced HF. Selective β3AR stimulation had a positive inotropic effect. Transgenic mice constitutively overexpressing human β3AR in the heart (c-hβ3tg) were protected from the development of HF in response to induced AS, and against cardiomyocyte mitochondrial dysfunction (fragmented mitochondria with remodeled cristae and metabolic reprogramming featuring altered substrate use). Similar beneficial effects were observed in wild-type mice inoculated with adeno-associated virus (AAV9) inducing cardiac-specific overexpression of human β3AR before AS induction. Moreover, AAV9-hβ3AR injection into wild-type mice at late disease stages, when cardiac hypertrophy and metabolic reprogramming are already advanced, reversed the HF phenotype and restored balanced mitochondrial dynamics, demonstrating the potential of gene-therapy-mediated β3AR overexpression in AS. Mice with cardiac specific ablation of Yme1l (cYKO), characterized by fragmented mitochondria, showed an increased mortality upon AS challenge. AAV9-hβ3AR injection in these mice before AS induction reverted the fragmented mitochondria phenotype and rescued them from death. In conclusion, our results step out that β3AR overexpression might have translational potential as a therapeutic strategy in AS-induced HF.Sin financiación12.416 JCR (2021) Q1, 13/143 Cardiac & Cardiovascular Systems1.615 SJR (2021) Q1, 49/356 Cardiology and Cardiovascular MedicineNo data IDR 2021UE

Neutrophil β1 adrenergic receptor blockade blunts stroke-associated neuroinflammation

39 p.-7 fig.-1 graph. abst.Background and Purpose Reperfusion therapy is the standard of care for ischemic stroke; however, there is a need to identify new therapeutic targets able to ameliorate cerebral damage. Neutrophil´s β1 adrenergic receptor (β1AR) has been linked to migration during exacerbated inflammation. Given the central role of neutrophils in cerebral damage during stroke, we hypothesize that β1AR blockade will improve stroke outcomes.Experimental Approach Rats were subjected to middle cerebral artery occlusion–reperfusion to evaluate the effect on stroke of the selective β1AR blocker metoprolol (12.5 mg/Kg) when injected i.v. 10 min before reperfusion.Key Results Magnetic resonance imaging and histopathology analysis showed that pre-reperfusion i.v. metoprolol reduced IS. This was accompanied by reduced cytotoxic edema at 24 h and vasogenic edema at 7 d. Metoprolol-treated rats showed reduced brain neutrophil infiltration and those which infiltrated displayed a high proportion of anti-inflammatory phenotype (N2, YM1+). Additional inflammatory models demonstrated that metoprolol specifically blocked neutrophil migration via β1AR and excluded a significant effect on glia compartment. Consistently, metoprolol did not protect the brain in neutrophil-depleted rats upon stroke. In patients suffering an ischemic stroke, β1AR blockade by metoprolol reduced circulating neutrophil–platelet co-aggregates.Conclusions and Implications Our findings describe that β1AR blockade ameliorates cerebral damage by targeting neutrophils, identifying a novel therapeutic target to improve outcomes in patients with stroke. This therapeutic strategy is in the earliest stages of the translational pathway and should be further explored.This study received funding from the Instituto de Salud Carlos III (ISCIII; PI16/02110 to B.I and PT20/00044 to MD), the European Regional Development Fund (ERDF) “A way of making Europe", the Comunidad de Madrid (S2017/BMD-3867 RENIM-CM to MD and BI) cofunded with European structural and investment funds and by Agencia Estatal de Investigación (PID2019‐110369RB‐I00 to B.I). BI is a recipient of funding from the European Research Council (ERC) under the European Union Horizon 2020 Research and Innovation Programme (ERC-Consolidator Grant agreement No. 819775). EO is a recipient of funds from the Comunidad de Madrid Programa de Atracción de Talento (2017-T1/BMD 5185) and from a Ramón y Cajal grant (RYC2020-028884-I) funded by MCIN/AEI/10.13039/501100011033 and by “ESF Investing in your future”. ACM is the beneficiary of an FPU fellowship from the Ministerio de Ciencia e Innovación (FPU2017/01932). MCC is the beneficiary of a Miguel Servet contract (MS16/00174). The CNIC is supported by the ISCIII, the Ministerio de Ciencia e Innovación and the Pro CNIC Foundation and is a Severo Ochoa Center of Excellence (CEX2020-001041-S).Peer reviewe

Beta-3 adrenergic receptor overexpression reverses aortic stenosis-induced heart failure and restores balanced mitochondrial dynamics.

Aortic stenosis (AS) is associated with left ventricular (LV) hypertrophy and heart failure (HF). There is a lack of therapies able to prevent/revert AS-induced HF. Beta3 adrenergic receptor (β3AR) signaling is beneficial in several forms of HF. Here, we studied the potential beneficial effect of β3AR overexpression on AS-induced HF. Selective β3AR stimulation had a positive inotropic effect. Transgenic mice constitutively overexpressing human β3AR in the heart (c-hβ3tg) were protected from the development of HF in response to induced AS, and against cardiomyocyte mitochondrial dysfunction (fragmented mitochondria with remodeled cristae and metabolic reprogramming featuring altered substrate use). Similar beneficial effects were observed in wild-type mice inoculated with adeno-associated virus (AAV9) inducing cardiac-specific overexpression of human β3AR before AS induction. Moreover, AAV9-hβ3AR injection into wild-type mice at late disease stages, when cardiac hypertrophy and metabolic reprogramming are already advanced, reversed the HF phenotype and restored balanced mitochondrial dynamics, demonstrating the potential of gene-therapy-mediated β3AR overexpression in AS. Mice with cardiac specific ablation of Yme1l (cYKO), characterized by fragmented mitochondria, showed an increased mortality upon AS challenge. AAV9-hβ3AR injection in these mice before AS induction reverted the fragmented mitochondria phenotype and rescued them from death. In conclusion, our results step out that β3AR overexpression might have translational potential as a therapeutic strategy in AS-induced HF.Spanish Ministry of Science and Innovation (MICINN) RETOS2019-107332RB-I00, European Commission (ERC-CoG grant N° 819775), ERA-CVD Joint Translational Call 2016 (funded through the Instituto de Salud Carlos III (ISCIII) and the European Regional Development Fund (ERDF), # AC16/00021), and BBVA foundation grant (# BIO CAR 0265) to B.I. PID2019-104776RB-I00, CB16/11/00399 (CIBER CV), and RD16/0011/0021 (TERCEL) from MCIN/AEI/ https://doi.org/10.13039/501100011033and a grant from the Fundación BBVA (Ref.: BIO14_298) to J.L.d.l.P. E.O. is recipient of funds from Programa de Atracción de Talento (2017-T1/BMD-5185) of Comunidad de Madrid and from a Ramón y Cajal grant (RYC2020- 028884-I) funded by MCIN/AEI/ https://doi.org/10.13039/50110 0011033 and by “ESF Investing in your future. A.P. is benefciary of a FPI fellowship by the MCI (BES-2012–061091), R.V-G. of a Spanish National doctorate fellowship funded by ISCIII (Contratos PFIS FI17/00045) and A.C-M. is benefciary of a FPU fellowship from the MCI (FPU2017/01932). This study was partially supported by the Comunidad de Madrid (RENIM-CM, S2017/BMD-3867 & P2022/ BMD-7403) and cofunded with European structural and investment funds. The CNIC is supported by the ISCIII, the MICINN and the Pro CNIC Foundation, and is a Center of Excellence Severo Ochoa (grant CEX2020-001041-S funded by MICIN/AEI/https://doi.org/10.13039/ 501100011033).S