Cardiomyocyte-Targeting Peptide to Deliver Amiodarone
Background: Amiodarone is underutilized due to significant off-target toxicities. We hypothesized that targeted delivery to the heart would lead to the lowering of the dose by utilizing a cardiomyocyte-targeting peptide (CTP), a cell-penetrating peptide identified by our prior phage display work. Me...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-08-01
|
| Series: | Pharmaceutics |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1999-4923/15/8/2107 |
| _version_ | 1827728901471731712 |
|---|---|
| author | Maliha Zahid Beth Weber Ray Yurko Kazi Islam Vaishavi Agrawal Jack Lopuszynski Hisato Yagi Guy Salama |
| author_facet | Maliha Zahid Beth Weber Ray Yurko Kazi Islam Vaishavi Agrawal Jack Lopuszynski Hisato Yagi Guy Salama |
| author_sort | Maliha Zahid |
| collection | DOAJ |
| description | Background: Amiodarone is underutilized due to significant off-target toxicities. We hypothesized that targeted delivery to the heart would lead to the lowering of the dose by utilizing a cardiomyocyte-targeting peptide (CTP), a cell-penetrating peptide identified by our prior phage display work. Methods: CTP was synthesized thiolated at the N-terminus, conjugated to amiodarone via Schiff base chemistry, HPLC purified, and confirmed with MALDI/TOF. The stability of the conjugate was assessed using serial HPLCs. Guinea pigs (GP) were injected intraperitoneally daily with vehicle (7 days), amiodarone (7 days; 80 mg/kg), CTP–amiodarone (5 days; 26.3 mg/kg), or CTP (5 days; 17.8 mg/kg), after which the GPs were euthanized, and the hearts were excised and perfused on a Langendorff apparatus with Tyrode’s solution and blebbistatin (5 µM) to minimize the contractions. Voltage (RH237) and Ca<sup>2+</sup>-indicator dye (Rhod-2/AM) were injected, and fluorescence from the epicardium split and was captured by two cameras at 570–595 nm for the cytosolic Ca<sup>2+</sup> and 610–750 nm wavelengths for the voltage. Subsequently, the hearts were paced at 250 ms with programmed stimulation to measure the changes in the conduction velocities (CV), action potential duration (APD), and Ca<sup>2+</sup> transient durations at 90% recovery (CaTD<sub>90</sub>). mRNA was extracted from all hearts, and RNA sequencing was performed with results compared to the control hearts. Results: The CTP–amiodarone remained stable for up to 21 days at 37 °C. At ~1/15th of the dose of amiodarone, the CTP–amiodarone decreased the CV in hearts significantly compared to the control GPs (0.92 ± 0.05 vs. 1.00 ± 0.03 ms, <i>p</i> = 0.0007), equivalent to amiodarone alone (0.87 ± 0.08 ms, <i>p</i> = 0.0003). Amiodarone increased the APD (192 ± 5 ms vs. 175 ± 8 ms for vehicle, <i>p</i> = 0.0025), while CTP–amiodarone decreased it significantly (157 ± 16 ms, <i>p</i> = 0.0136), similar to CTP alone (155 ± 13 ms, <i>p</i> = 0.0039). Both amiodarone and CTP–amiodarone significantly decreased the calcium transients compared to the controls. CTP–amiodarone and CTP decreased the CaTD<sub>90</sub> to an extent greater than amiodarone alone (<i>p</i> < 0.001). RNA-seq showed that CTP alone increased the expression of DHPR and SERCA2a, while it decreased the expression of the proinflammatory genes, NF-kappa B, TNF-α, IL-1β, and IL-6. Conclusions: Our data suggest that CTP can deliver amiodarone to cardiomyocytes at ~1/15th the total molar dose of the amiodarone needed to produce a comparable slowing of CVs. The ability of CTP to decrease the AP durations and CaTD<sub>90</sub> may be related to its increase in the expression of Ca-handling genes, which merits further study. |
| first_indexed | 2024-03-10T23:39:26Z |
| format | Article |
| id | doaj.art-4715d0ab7cca4914b0c2ebd9d5b30beb |
| institution | Directory Open Access Journal |
| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-10T23:39:26Z |
| publishDate | 2023-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceutics |
| spelling | doaj.art-4715d0ab7cca4914b0c2ebd9d5b30beb2023-11-19T02:36:55ZengMDPI AGPharmaceutics1999-49232023-08-01158210710.3390/pharmaceutics15082107Cardiomyocyte-Targeting Peptide to Deliver AmiodaroneMaliha Zahid0Beth Weber1Ray Yurko2Kazi Islam3Vaishavi Agrawal4Jack Lopuszynski5Hisato Yagi6Guy Salama7Department of Cardiovascular Diseases, Mayo Clinic, Rochester, MN 55905, USAPittsburgh Heart, Lung, Blood, and Vascular Medicine Institute, Division of Cardiology, Department of Medicine, University of Pittsburgh School of Medicine, University of Pittsburgh Medical Center, Pittsburgh, PA 15213, USAPeptide Synthesis Facility, University of Pittsburgh, Pittsburgh, PA 15219, USAPeptide Synthesis Facility, University of Pittsburgh, Pittsburgh, PA 15219, USADietrich School of Arts and Sciences, University of Pittsburgh, Pittsburgh, PA 15260, USADepartment of Cardiovascular Diseases, Mayo Clinic, Rochester, MN 55905, USADepartment of Developmental Biology, University of Pittsburgh, Pittsburgh, PA 15201, USAPittsburgh Heart, Lung, Blood, and Vascular Medicine Institute, Division of Cardiology, Department of Medicine, University of Pittsburgh School of Medicine, University of Pittsburgh Medical Center, Pittsburgh, PA 15213, USABackground: Amiodarone is underutilized due to significant off-target toxicities. We hypothesized that targeted delivery to the heart would lead to the lowering of the dose by utilizing a cardiomyocyte-targeting peptide (CTP), a cell-penetrating peptide identified by our prior phage display work. Methods: CTP was synthesized thiolated at the N-terminus, conjugated to amiodarone via Schiff base chemistry, HPLC purified, and confirmed with MALDI/TOF. The stability of the conjugate was assessed using serial HPLCs. Guinea pigs (GP) were injected intraperitoneally daily with vehicle (7 days), amiodarone (7 days; 80 mg/kg), CTP–amiodarone (5 days; 26.3 mg/kg), or CTP (5 days; 17.8 mg/kg), after which the GPs were euthanized, and the hearts were excised and perfused on a Langendorff apparatus with Tyrode’s solution and blebbistatin (5 µM) to minimize the contractions. Voltage (RH237) and Ca<sup>2+</sup>-indicator dye (Rhod-2/AM) were injected, and fluorescence from the epicardium split and was captured by two cameras at 570–595 nm for the cytosolic Ca<sup>2+</sup> and 610–750 nm wavelengths for the voltage. Subsequently, the hearts were paced at 250 ms with programmed stimulation to measure the changes in the conduction velocities (CV), action potential duration (APD), and Ca<sup>2+</sup> transient durations at 90% recovery (CaTD<sub>90</sub>). mRNA was extracted from all hearts, and RNA sequencing was performed with results compared to the control hearts. Results: The CTP–amiodarone remained stable for up to 21 days at 37 °C. At ~1/15th of the dose of amiodarone, the CTP–amiodarone decreased the CV in hearts significantly compared to the control GPs (0.92 ± 0.05 vs. 1.00 ± 0.03 ms, <i>p</i> = 0.0007), equivalent to amiodarone alone (0.87 ± 0.08 ms, <i>p</i> = 0.0003). Amiodarone increased the APD (192 ± 5 ms vs. 175 ± 8 ms for vehicle, <i>p</i> = 0.0025), while CTP–amiodarone decreased it significantly (157 ± 16 ms, <i>p</i> = 0.0136), similar to CTP alone (155 ± 13 ms, <i>p</i> = 0.0039). Both amiodarone and CTP–amiodarone significantly decreased the calcium transients compared to the controls. CTP–amiodarone and CTP decreased the CaTD<sub>90</sub> to an extent greater than amiodarone alone (<i>p</i> < 0.001). RNA-seq showed that CTP alone increased the expression of DHPR and SERCA2a, while it decreased the expression of the proinflammatory genes, NF-kappa B, TNF-α, IL-1β, and IL-6. Conclusions: Our data suggest that CTP can deliver amiodarone to cardiomyocytes at ~1/15th the total molar dose of the amiodarone needed to produce a comparable slowing of CVs. The ability of CTP to decrease the AP durations and CaTD<sub>90</sub> may be related to its increase in the expression of Ca-handling genes, which merits further study.https://www.mdpi.com/1999-4923/15/8/2107cardiac targeting peptidecell penetrating peptidesamiodaroneatrial fibrillation |
| spellingShingle | Maliha Zahid Beth Weber Ray Yurko Kazi Islam Vaishavi Agrawal Jack Lopuszynski Hisato Yagi Guy Salama Cardiomyocyte-Targeting Peptide to Deliver Amiodarone Pharmaceutics cardiac targeting peptide cell penetrating peptides amiodarone atrial fibrillation |
| title | Cardiomyocyte-Targeting Peptide to Deliver Amiodarone |
| title_full | Cardiomyocyte-Targeting Peptide to Deliver Amiodarone |
| title_fullStr | Cardiomyocyte-Targeting Peptide to Deliver Amiodarone |
| title_full_unstemmed | Cardiomyocyte-Targeting Peptide to Deliver Amiodarone |
| title_short | Cardiomyocyte-Targeting Peptide to Deliver Amiodarone |
| title_sort | cardiomyocyte targeting peptide to deliver amiodarone |
| topic | cardiac targeting peptide cell penetrating peptides amiodarone atrial fibrillation |
| url | https://www.mdpi.com/1999-4923/15/8/2107 |
| work_keys_str_mv | AT malihazahid cardiomyocytetargetingpeptidetodeliveramiodarone AT bethweber cardiomyocytetargetingpeptidetodeliveramiodarone AT rayyurko cardiomyocytetargetingpeptidetodeliveramiodarone AT kaziislam cardiomyocytetargetingpeptidetodeliveramiodarone AT vaishaviagrawal cardiomyocytetargetingpeptidetodeliveramiodarone AT jacklopuszynski cardiomyocytetargetingpeptidetodeliveramiodarone AT hisatoyagi cardiomyocytetargetingpeptidetodeliveramiodarone AT guysalama cardiomyocytetargetingpeptidetodeliveramiodarone |