Development of conditional-siRNA programmable riboswitch for targeting adverse cardiac remodeling
Heart failure (HF) remains a significant healthcare burden, with an unmet need for novel therapies to target the preceding pathological hypertrophy in HF patients. Here we report the development of novel conditional-siRNA (Cond-siRNA) constructs that are selectively activated by disease-specific RNA...
| Published in: | Molecular Therapy: Nucleic Acids |
|---|---|
| Main Authors: | , , , , , , , , , , , , |
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-09-01
|
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253125002215 |
| _version_ | 1849420899648798720 |
|---|---|
| author | Priyanka Gokulnath Ane M. Salvador Caleb Graham Si-ping Han Guoping Li Ramaswamy Kannappan Christopher Azzam Michail Spanos Lisa Scherer Palaniappan Sethu John Rossi William A. Goddard, III Saumya Das |
| author_facet | Priyanka Gokulnath Ane M. Salvador Caleb Graham Si-ping Han Guoping Li Ramaswamy Kannappan Christopher Azzam Michail Spanos Lisa Scherer Palaniappan Sethu John Rossi William A. Goddard, III Saumya Das |
| author_sort | Priyanka Gokulnath |
| collection | DOAJ |
| container_title | Molecular Therapy: Nucleic Acids |
| description | Heart failure (HF) remains a significant healthcare burden, with an unmet need for novel therapies to target the preceding pathological hypertrophy in HF patients. Here we report the development of novel conditional-siRNA (Cond-siRNA) constructs that are selectively activated by disease-specific RNA biomarkers to enable cell-specific inhibition of a target disease-causing RNA. We designed a Cond-siRNA that can be activated by Nppa mRNA, upregulated specifically in cardiomyocytes (CMs) under pathological stress, to silence the key pro-hypertrophic gene calcineurin (CaN) A-a by the effector small interfering RNA (siRNA). In both neonatal rat ventricular myocytes (NRVMs) and H9c2 CMs, Cond-siRNA showed minimal baseline activity but selectively silenced CaN upon Nppa mRNA induction by phenylephrine (PE) stress in cell culture models and pressure overload (PO) in a heart-on-a-chip model. In NRVMs, Cond-siRNA reduced CaN mRNA only after PE or PO, but not with vehicle, confirming Nppa-specific activation. This specificity was further validated as Cond-siRNA did not affect CaN in cardiac fibroblasts or T cells lacking Nppa. Reduced CaN protein levels and NFATc1 nuclear translocation correlated with decreased NRVM hypertrophy after PE treatment, confirming Cond-siRNA’s efficacy. This study offers proof-of-concept for Cond-siRNA as a targeted therapy to mitigate hypertrophic progression, paving the way for novel HF treatments. |
| format | Article |
| id | doaj-art-eafa3149c8594c08b411209d1008ea9a |
| institution | Directory of Open Access Journals |
| issn | 2162-2531 |
| language | English |
| publishDate | 2025-09-01 |
| publisher | Elsevier |
| record_format | Article |
| spelling | doaj-art-eafa3149c8594c08b411209d1008ea9a2025-08-20T03:44:06ZengElsevierMolecular Therapy: Nucleic Acids2162-25312025-09-0136310266710.1016/j.omtn.2025.102667Development of conditional-siRNA programmable riboswitch for targeting adverse cardiac remodelingPriyanka Gokulnath0Ane M. Salvador1Caleb Graham2Si-ping Han3Guoping Li4Ramaswamy Kannappan5Christopher Azzam6Michail Spanos7Lisa Scherer8Palaniappan Sethu9John Rossi10William A. Goddard, III11Saumya Das12Cardiovascular Research Center, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USACardiovascular Research Center, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USAUniversity of Alabama at Birmingham, Birmingham, AL 35294, USADepartment of Molecular and Cellular Biology, City of Hope, Duarte, CA 91010, USACardiovascular Research Center, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USAUniversity of Alabama at Birmingham, Birmingham, AL 35294, USACardiovascular Research Center, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USACardiovascular Research Center, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USADepartment of Molecular and Cellular Biology, City of Hope, Duarte, CA 91010, USAUniversity of Alabama at Birmingham, Birmingham, AL 35294, USADepartment of Molecular and Cellular Biology, City of Hope, Duarte, CA 91010, USAMaterials and Process Simulation Center, California Institute of Technology, Pasadena, CA 91125, USACardiovascular Research Center, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA; Corresponding author: Saumya Das, MD, PhD, Professor of Medicine, Cardiology Division, Cardiovascular Research Center, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA.Heart failure (HF) remains a significant healthcare burden, with an unmet need for novel therapies to target the preceding pathological hypertrophy in HF patients. Here we report the development of novel conditional-siRNA (Cond-siRNA) constructs that are selectively activated by disease-specific RNA biomarkers to enable cell-specific inhibition of a target disease-causing RNA. We designed a Cond-siRNA that can be activated by Nppa mRNA, upregulated specifically in cardiomyocytes (CMs) under pathological stress, to silence the key pro-hypertrophic gene calcineurin (CaN) A-a by the effector small interfering RNA (siRNA). In both neonatal rat ventricular myocytes (NRVMs) and H9c2 CMs, Cond-siRNA showed minimal baseline activity but selectively silenced CaN upon Nppa mRNA induction by phenylephrine (PE) stress in cell culture models and pressure overload (PO) in a heart-on-a-chip model. In NRVMs, Cond-siRNA reduced CaN mRNA only after PE or PO, but not with vehicle, confirming Nppa-specific activation. This specificity was further validated as Cond-siRNA did not affect CaN in cardiac fibroblasts or T cells lacking Nppa. Reduced CaN protein levels and NFATc1 nuclear translocation correlated with decreased NRVM hypertrophy after PE treatment, confirming Cond-siRNA’s efficacy. This study offers proof-of-concept for Cond-siRNA as a targeted therapy to mitigate hypertrophic progression, paving the way for novel HF treatments.http://www.sciencedirect.com/science/article/pii/S2162253125002215MT: Oligonucleotides: Therapies and ApplicationssiRNARNA therapeuticscardiac hypertrophyheart-on-chip modelriboswitch |
| spellingShingle | Priyanka Gokulnath Ane M. Salvador Caleb Graham Si-ping Han Guoping Li Ramaswamy Kannappan Christopher Azzam Michail Spanos Lisa Scherer Palaniappan Sethu John Rossi William A. Goddard, III Saumya Das Development of conditional-siRNA programmable riboswitch for targeting adverse cardiac remodeling MT: Oligonucleotides: Therapies and Applications siRNA RNA therapeutics cardiac hypertrophy heart-on-chip model riboswitch |
| title | Development of conditional-siRNA programmable riboswitch for targeting adverse cardiac remodeling |
| title_full | Development of conditional-siRNA programmable riboswitch for targeting adverse cardiac remodeling |
| title_fullStr | Development of conditional-siRNA programmable riboswitch for targeting adverse cardiac remodeling |
| title_full_unstemmed | Development of conditional-siRNA programmable riboswitch for targeting adverse cardiac remodeling |
| title_short | Development of conditional-siRNA programmable riboswitch for targeting adverse cardiac remodeling |
| title_sort | development of conditional sirna programmable riboswitch for targeting adverse cardiac remodeling |
| topic | MT: Oligonucleotides: Therapies and Applications siRNA RNA therapeutics cardiac hypertrophy heart-on-chip model riboswitch |
| url | http://www.sciencedirect.com/science/article/pii/S2162253125002215 |
| work_keys_str_mv | AT priyankagokulnath developmentofconditionalsirnaprogrammableriboswitchfortargetingadversecardiacremodeling AT anemsalvador developmentofconditionalsirnaprogrammableriboswitchfortargetingadversecardiacremodeling AT calebgraham developmentofconditionalsirnaprogrammableriboswitchfortargetingadversecardiacremodeling AT sipinghan developmentofconditionalsirnaprogrammableriboswitchfortargetingadversecardiacremodeling AT guopingli developmentofconditionalsirnaprogrammableriboswitchfortargetingadversecardiacremodeling AT ramaswamykannappan developmentofconditionalsirnaprogrammableriboswitchfortargetingadversecardiacremodeling AT christopherazzam developmentofconditionalsirnaprogrammableriboswitchfortargetingadversecardiacremodeling AT michailspanos developmentofconditionalsirnaprogrammableriboswitchfortargetingadversecardiacremodeling AT lisascherer developmentofconditionalsirnaprogrammableriboswitchfortargetingadversecardiacremodeling AT palaniappansethu developmentofconditionalsirnaprogrammableriboswitchfortargetingadversecardiacremodeling AT johnrossi developmentofconditionalsirnaprogrammableriboswitchfortargetingadversecardiacremodeling AT williamagoddardiii developmentofconditionalsirnaprogrammableriboswitchfortargetingadversecardiacremodeling AT saumyadas developmentofconditionalsirnaprogrammableriboswitchfortargetingadversecardiacremodeling |