Optical modulation of excitation-contraction coupling in human-induced pluripotent stem cell-derived cardiomyocytes

Vurro, Vito; Federici, Beatrice; Ronchi, Carlotta; Florindi, Chiara; Sesti, Valentina; Crasto, Silvia; Maniezzi, Claudia; Galli, Camilla; Antognazza, Maria Rosa; Bertarelli, Chiara; Pasquale, Elisa Di; Lanzani, Guglielmo; Lodola, Francesco

doi:10.1016/j.isci.2023.106121

Cited by 9 publications

(14 citation statements)

References 61 publications

(73 reference statements)

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“…We have recently demonstrated that the photoactivation of an intramembrane photochromic transducer, Ziapin2, enhances both the frequency of Ca 2+ transients and the contraction rate of hPSC‐CMs. [ 59 ] However, the biophysical mechanism ruling the photostimulation of Ziapin2‐loaded hiPSC‐CMs is not yet fully elucidated, making a direct comparison with the present work not straightforward.…”

Section: Resultsmentioning

confidence: 99%

Nongenetic Optical Modulation of Pluripotent Stem Cells Derived Cardiomyocytes Function in the Red Spectral Range

Ronchi,

Galli,

Tullii

et al. 2023

Advanced Science

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Optical stimulation in the red/near infrared range recently gained increasing interest, as a not‐invasive tool to control cardiac cell activity and repair in disease conditions. Translation of this approach to therapy is hampered by scarce efficacy and selectivity. The use of smart biocompatible materials, capable to act as local, NIR‐sensitive interfaces with cardiac cells, may represent a valuable solution, capable to overcome these limitations. In this work, a far red‐responsive conjugated polymer, namely poly[2,1,3‐benzothiadiazole‐4,7‐diyl[4,4‐bis(2‐ethylhexyl)−4H‐cyclopenta[2,1‐b:3,4‐b’]dithiophene‐2,6‐diyl]] (PCPDTBT) is proposed for the realization of photoactive interfaces with cardiomyocytes derived from pluripotent stem cells (hPSC‐CMs). Optical excitation of the polymer turns into effective ionic and electrical modulation of hPSC‐CMs, in particular by fastening Ca2+ dynamics, inducing action potential shortening, accelerating the spontaneous beating frequency. The involvement in the phototransduction pathway of Sarco‐Endoplasmic Reticulum Calcium ATPase (SERCA) and Na+/Ca2+ exchanger (NCX) is proven by pharmacological assays and is correlated with physical/chemical processes occurring at the polymer surface upon photoexcitation. Very interestingly, an antiarrhythmogenic effect, unequivocally triggered by polymer photoexcitation, is also observed. Overall, red‐light excitation of conjugated polymers may represent an unprecedented opportunity for fine control of hPSC‐CMs functionality and can be considered as a perspective, noninvasive approach to treat arrhythmias.

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Section: Resultsmentioning

confidence: 99%

Nongenetic Optical Modulation of Pluripotent Stem Cells Derived Cardiomyocytes Function in the Red Spectral Range

Ronchi,

Galli,

Tullii

et al. 2023

Advanced Science

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“…This phenomenon has been reported in a variety of different types of cells, including bacteria, HEK293T cells, neurons, and cardiomyocytes. In excitable cells, the membrane potential variation is sufficient to trigger action potentials. ,,, …”

Section: Resultsmentioning

confidence: 99%

“…In excitable cells, the membrane potential variation is sufficient to trigger action potentials. 47,48,51,52 Since Ziapin2 dimerization generates a light reversible thinning of the membrane, we recorded cell membrane capacitance of both Ctrl and hTRAAK cells before and during visible light illumination to reduce the impact of cell intrinsic variability. Cell membrane capacitance was assessed by applying a voltage step (ΔV) of 5 mV.…”

Section: ■ Resultsmentioning

confidence: 99%

“…The insertion in the membrane and the consequent formation of Ziapin2 dimers lead to a thinning of the membrane and an increase of the cell capacitance . Visible light pulses induce trans -to- cis isomerization that leads to the breaking of Ziapin2 dimers, with a significant perturbation of the cell membrane potentials. , Specifically, during illumination, membrane relaxation generates a rapid hyperpolarization followed by slight depolarization after the end of the stimulus. − In the present work, we expressed human TRAAK (hTRAAK) channels into HEK293T (Human Embryonic Kidney) cells via transfection, observing that Ziapin2 insertion in the membrane is able per se to recruit part of the hTRAAK channels permitting the exit of K + ions outside the cells. K + outward flux generates a sustained increase in the current and consequently a 2-fold hyperpolarization of the cell membrane.…”

Section: Introductionmentioning

confidence: 99%

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Modulation of Mechanosensitive Potassium Channels by a Membrane-targeted Nongenetic Photoswitch

Moschetta,

Vurro,

Sesti

et al. 2023

J. Phys. Chem. B

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Mechanosensitive ion channels are present in the plasma membranes of all cells. They play a fundamental role in converting mechanical stimuli into biochemical signals and are involved in several physiological processes such as touch sensation, hearing, and blood pressure regulation. This protein family includes TWIK-related arachidonic acid-stimulated K+ channel (TRAAK), which is specifically implicated in the maintenance of the resting membrane potential and in the regulation of a variety of important neurobiological functions. Dysregulation of these channels has been linked to various diseases, including blindness, epilepsy, cardiac arrhythmia, and chronic pain. For these reasons, mechanosensitive channels are targets for the treatment of several diseases. Here, we propose a new approach to investigate TRAAK ion channel modulation that is based on nongenetic photostimulation. We employed an amphiphilic azobenzene, named Ziapin2. In the dark, Ziapin2 preferentially dwells in the plasma membrane, causing a thinning of the membrane. Upon light irradiation, an isomerization occurs, breaking the dimers and inducing membrane relaxation. To study the effect of Ziapin2 on the mechanosensitive channels, we expressed human TRAAK (hTRAAK) channels in HEK293T cells. We observed that Ziapin2 insertion in the membrane is able per se to recruit hTRAAK, permitting the exit of K+ ions outside the cells with a consequent hyperpolarization of the cell membrane. During light stimulation, membrane relaxation induces hTRAAK closure, generating a consistent and compensatory depolarization. These results add information to the Ziapin2 mechanism and suggest that membrane deformation can be a tool for the nonselective modulation of mechanosensitive channels.

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“…Apart from optogenetics, an alternative photostimulation paradigm relies on exogenous light-responsive materials, usually in molecular or nanostructured forms [ 8 , 9 , 10 ]. In particular, we here refer to a family of amphiphilic photochromic azobenzenes, recently demonstrated to be molecular transducers [ 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 ]. Optical modulation of cell parameters stems from the excitation of the azobenzene that in turn releases energy to the membrane, possibly changing the membrane thickness [ 19 ].…”

Section: Introductionmentioning

confidence: 99%

Membrane Order Effect on the Photoresponse of an Organic Transducer

et al. 2023

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Non-genetic photostimulation, which allows for control over cellular activity via the use of cell-targeting phototransducers, is widely used nowadays to study and modulate/restore biological functions. This approach relies on non-covalent interactions between the phototransducer and the cell membrane, thus implying that cell conditions and membrane status can dictate the effectiveness of the method. For instance, although immortalized cell lines are traditionally used in photostimulation experiments, it has been demonstrated that the number of passages they undergo is correlated to the worsening of cell conditions. In principle, this could impact cell responsivity against exogenous stressors, including photostimulation. However, these aspects have usually been neglected in previous experiments. In this work, we investigated whether cell passages could affect membrane properties (such as polarity and fluidity). We applied optical spectroscopy and electrophysiological measurements in two different biological models: (i) an epithelial immortalized cell line (HEK-293T cells) and (ii) liposomes. Different numbers of cell passages were compared to a different morphology in the liposome membrane. We demonstrated that cell membranes show a significant decrease in ordered domains upon increasing the passage number. Furthermore, we observed that cell responsivity against external stressors is markedly different between aged and non-aged cells. Firstly, we noted that the thermal-disordering effect that is usually observed in membranes is more evident in aged cells than in non-aged ones. We then set up a photostimulation experiment by using a membrane-targeted azobenzene as a phototransducer (Ziapin2). As an example of a functional consequence of such a condition, we showed that the rate of isomerization of an intramembrane molecular transducer is significantly impaired in aged cells. The reduction in the photoisomerization rate translates in cells with a sustained reduction of the Ziapin2-related hyperpolarization of the membrane potential and an overall increase in the molecule fluorescence. Overall, our results suggest that membrane stimulation strongly depends on membrane order, highlighting the importance of cell passage during the characterization of the stimulation tools. This study can shine light on the correlation between aging and the development of diseases driven by membrane degradation as well as on the different cell responsivities against external stressors, such as temperature and photostimulation.

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Optical modulation of excitation-contraction coupling in human-induced pluripotent stem cell-derived cardiomyocytes

Cited by 9 publications

References 61 publications

Nongenetic Optical Modulation of Pluripotent Stem Cells Derived Cardiomyocytes Function in the Red Spectral Range

Nongenetic Optical Modulation of Pluripotent Stem Cells Derived Cardiomyocytes Function in the Red Spectral Range

Modulation of Mechanosensitive Potassium Channels by a Membrane-targeted Nongenetic Photoswitch

Membrane Order Effect on the Photoresponse of an Organic Transducer

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