Lipoplexes for effective in vitro delivery of microRNAs to adult human cardiac fibroblasts for perspective direct cardiac cell reprogramming

Nicoletti, Letizia; Paoletti, Camilla; Tarricone, Giulia; Andreana, Ilaria; Stella, Barbara; Berlier, Gloria; Divieto, Carla; Mattu, Clara; Chiono, Valeria

doi:10.1016/j.nano.2022.102589

Cited by 10 publications

(16 citation statements)

References 46 publications

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“…Furthermore, this approach was tested injecting this nanocomplex into the epicardium along the border zone of infarcted mice, which, after 4 weeks, showed a significant decrease in cardiac fibrosis and infarct thickness [ 77 ]. Recently, Nicoletti and colleagues investigated a lipoplex based nanosystem to carry miRcombo in adult human CFs [ 78 ]. In this study, lipoplexes were composed of helper dioleoyl phosphatidylethanolamine (DOPE) and cationic lipid [2-(2,3-didodecyloxypropyl)-hydroxyethyl] ammonium bromide (DE).…”

Section: Cf Reprogramming Into Induced Cmsmentioning

confidence: 99%

“…In an in vitro study, DE-DOPE lipoplexes with nanometric hydrodynamic size (372 nm) showed high loading proficiency (99%), quicker miRNA release (99%), and high viability (80–100%) compared to commercial lipoplexes. In addition, DE-DOPE lipoplexes increased CF reprogramming into iCM cells as indicated by increased expression of CM markers [ 78 ].…”

Section: Cf Reprogramming Into Induced Cmsmentioning

confidence: 99%

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Direct Reprogramming of Resident Non-Myocyte Cells and Its Potential for In Vivo Cardiac Regeneration

Perveen

Vanni

Iacono

et al. 2023

Cells

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Cardiac diseases are the foremost cause of morbidity and mortality worldwide. The heart has limited regenerative potential; therefore, lost cardiac tissue cannot be replenished after cardiac injury. Conventional therapies are unable to restore functional cardiac tissue. In recent decades, much attention has been paid to regenerative medicine to overcome this issue. Direct reprogramming is a promising therapeutic approach in regenerative cardiac medicine that has the potential to provide in situ cardiac regeneration. It consists of direct cell fate conversion of one cell type into another, avoiding transition through an intermediary pluripotent state. In injured cardiac tissue, this strategy directs transdifferentiation of resident non-myocyte cells (NMCs) into mature functional cardiac cells that help to restore the native tissue. Over the years, developments in reprogramming methods have suggested that regulation of several intrinsic factors in NMCs can help to achieve in situ direct cardiac reprogramming. Among NMCs, endogenous cardiac fibroblasts have been studied for their potential to be directly reprogrammed into both induced cardiomyocytes and induced cardiac progenitor cells, while pericytes can transdifferentiate towards endothelial cells and smooth muscle cells. This strategy has been indicated to improve heart function and reduce fibrosis after cardiac injury in preclinical models. This review summarizes the recent updates and progress in direct cardiac reprogramming of resident NMCs for in situ cardiac regeneration.

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Section: Cf Reprogramming Into Induced Cmsmentioning

confidence: 99%

Section: Cf Reprogramming Into Induced Cmsmentioning

confidence: 99%

Direct Reprogramming of Resident Non-Myocyte Cells and Its Potential for In Vivo Cardiac Regeneration

Perveen

Vanni

Iacono

et al. 2023

Cells

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“…To this aim, a previously reported combination of four microRNAs, [30,32] called miRcombo (miR-1, miR-133, miR-208, and miR-499) was delivered to AHCFs by recently developed lipoplexes. [33] Two complementary experiments were performed with the aim to predict in vivo direct reprogramming outputs, elucidating: i) the capability of the therapy to prevent fibrotic traits in a mechanically active environment (i.e., cyclic mechanical stimulation applied to miRcombo transfected cell); ii) the potential to use the therapy to revert fibrotic traits (i.e., reprogramming performed in situ in a mechanically induced 3D cardiac fibrotic microenvironment). In both cases, the efficacy of miRcombo therapy was hampered by the mechanical stimulation, but not by the static 3D environment, suggesting the importance of screening therapies in relevant tissue models, recapitulating both 3D architecture and mechanical microenvironment.…”

Section: Introductionmentioning

confidence: 99%

In Vitro Mechanical Stimulation to Reproduce the Pathological Hallmarks of Human Cardiac Fibrosis on a Beating Chip and Predict The Efficacy of Drugs and Advanced Therapies

Visone,

Paoletti,

Cordiale

et al. 2023

Adv Healthcare Materials

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Cardiac fibrosis is one of the main causes of heart failure, significantly contributing to mortality. The discovery and development of effective therapies able to heal fibrotic pathological symptoms thus remain of paramount importance. Micro‐physiological systems (MPS) are recently introduced as promising platforms able to accelerate this finding. Here a 3D in vitro model of human cardiac fibrosis, named uScar, is developed by imposing a cyclic mechanical stimulation to human atrial cardiac fibroblasts (AHCFs) cultured in a 3D beating heart‐on‐chip and exploited to screen drugs and advanced therapeutics. The sole provision of a cyclic 10% uniaxial strain at 1 Hz to the microtissues is sufficient to trigger fibrotic traits, inducing a consistent fibroblast‐to‐myofibroblast transition and an enhanced expression and production of extracellular matrix (ECM) proteins. Standard of care anti‐fibrotic drugs (i.e., Pirfenidone and Tranilast) are confirmed to be efficient in preventing the onset of fibrotic traits in uScar. Conversely, the mechanical stimulation applied to the microtissues limit the ability of a miRNA therapy to directly reprogram fibroblasts into cardiomyocytes (CMs), despite its proved efficacy in 2D models. Such results demonstrate the importance of incorporating in vivo‐like stimulations to generate more representative 3D in vitro models able to predict the efficacy of therapies in patients.

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Section: Introductionmentioning

confidence: 99%

“…Hence, effective therapies for cardiac fibrosis regression and heart functional recovery are highly demanded. Advanced approaches for cardiac regeneration are under study and include cell reprogramming strategies 15,16 aimed at inducing cardiomyocyte proliferation or the trans-differentiation of CFs into cardiomyocytes, and stem cell therapies. 17 Preclinical validation of new advanced approaches/drugs could benefit from reliable in vitro models of human cardiac tissue, in agreement with the 3Rs principle (Reduction, Replacement, and Refinement).…”

Section: Introductionmentioning

confidence: 99%

Biomimetic Electrospun Scaffold-Based In Vitro Model Resembling the Hallmarks of Human Myocardial Fibrotic Tissue

Ruocco

Zoso

Mortati

et al. 2023

ACS Biomater. Sci. Eng.

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Adverse remodeling post-myocardial infarction is hallmarked by the phenotypic change of cardiac fibroblasts (CFs) into myofibroblasts (MyoFs) and over-deposition of the fibrotic extracellular matrix (ECM) mainly composed by fibronectin and collagens, with the loss of tissue anisotropy and tissue stiffening. Reversing cardiac fibrosis represents a key challenge in cardiac regenerative medicine. Reliable in vitro models of human cardiac fibrotic tissue could be useful for preclinical testing of new advanced therapies, addressing the limited predictivity of traditional 2D cell cultures and animal in vivo models. In this work, we engineered a biomimetic in vitro model, reproducing the morphological, mechanical, and chemical cues of native cardiac fibrotic tissue. Polycaprolactone (PCL)-based scaffolds with randomly oriented fibers were fabricated by solution electrospinning technique, showing homogeneous nanofibers with an average size of 131 ± 39 nm. PCL scaffolds were then surface-functionalized with human type I collagen (C1) and fibronectin (F) by dihydroxyphenylalanine (DOPA)-mediated mussel-inspired approach (PCL/polyDOPA/C1F), in order to mimic fibrotic cardiac tissue-like ECM composition and support human CF culture. BCA assay confirmed the successful deposition of the biomimetic coating and its stability during 5 days of incubation in phosphate-buffered saline. Immunostaining for C1 and F demonstrated their homogeneous distribution in the coating. AFM mechanical characterization showed that PCL/polyDOPA/C1F scaffolds, in wet conditions, resembled fibrotic tissue stiffness with an average Young's modulus of about 50 kPa. PCL/polyDOPA/C1F membranes supported human CF (HCF) adhesion and proliferation. Immunostaining for α-SMA and quantification of α-SMA-positive cells showed HCF activation into MyoFs in the absence of a transforming growth factor β (TGF-β) profibrotic stimulus, suggesting the intrinsic ability of biomimetic PCL/polyDOPA/C1F scaffolds to sustain the development of cardiac fibrotic tissue. A proof-ofconcept study making use of a commercially available antifibrotic drug confirmed the potentialities of the developed in vitro model for drug efficacy testing. In conclusion, the proposed model was able to replicate the main hallmarks of early-stage cardiac fibrosis, appearing as a promising tool for future preclinical testing of advanced regenerative therapies.

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Lipoplexes for effective in vitro delivery of microRNAs to adult human cardiac fibroblasts for perspective direct cardiac cell reprogramming

Cited by 10 publications

References 46 publications

Direct Reprogramming of Resident Non-Myocyte Cells and Its Potential for In Vivo Cardiac Regeneration

Direct Reprogramming of Resident Non-Myocyte Cells and Its Potential for In Vivo Cardiac Regeneration

In Vitro Mechanical Stimulation to Reproduce the Pathological Hallmarks of Human Cardiac Fibrosis on a Beating Chip and Predict The Efficacy of Drugs and Advanced Therapies

Biomimetic Electrospun Scaffold-Based In Vitro Model Resembling the Hallmarks of Human Myocardial Fibrotic Tissue

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