An electrical pulse can end in milliseconds. The cell that receives it may spend hours changing which parts of its genome it reads. Under certain experimental conditions, those changes can outlast the original signal.
A neuron fires. A muscle fiber contracts. Cells at the edge of a wound experience changes in their electrical environment. Each event can begin with ions moving across cell membranes, and each can start a chain of biochemical events that reaches the nucleus. There, the signal can influence transcription, change how accessible a region of DNA is, or alter chemical marks on DNA and the histones around which it is wrapped.
This is the scientific premise behind Lionheart Health’s research into programmable bioelectric signaling. Can a defined electrical pattern engage a pathway that changes the production or release of a specific protein? Can it coordinate several pathways involved in repair? And under what conditions do the target cells retain a measurable change after stimulation ends? Those questions connect electrical engineering to epigenetics, and they help explain why targets such as Klotho, SIRT6, BDNF, follistatin, SDF-1, COL17A1, and tropoelastin occupy different places in the same research program. (investors.lionheartlongevity.
1. Calcium carries the electrical signal to the nucleus
When membrane voltage changes, voltage-sensitive calcium channels can open. Calcium entering the cell binds to proteins such as calmodulin, which relays the signal to enzymes and transcription factors. The route calcium takes, where it enters, and how long it stays elevated all affect what reaches the nucleus.
One example is CREB, a transcription factor involved in activity-dependent gene expression. In hippocampal neurons, researchers found that the duration of stimulation helped determine how long CREB remained phosphorylated and how strongly it influenced transcription. In a separate experimental system, researchers showed that changing the frequency of calcium oscillations could change which transcriptional responses were activated. These findings explain why pulse width, burst spacing, and recovery intervals may matter alongside amplitude and frequency. They also explain why the pulse frequency displayed on a stimulator and the calcium pattern inside a cell need to be measured separately. [1–3] (web.stanford.edu, pubmed.ncbi.nlm.nih.gov, nature.com)
2. Histones and enhancers help determine which genes respond
DNA is wrapped around histone proteins. Chemical modifications to those histones can influence how readily a cell uses nearby regulatory DNA. One link between calcium signaling and this regulatory machinery is CREB-binding protein, or CBP. CBP helps activate transcription and can add acetyl groups to histones. In neurons, the calcium-sensitive kinase CaMKIV was shown to phosphorylate CBP and help drive activity-dependent transcription. [4] (pubmed.ncbi.nlm.nih.gov)
The next layer involves enhancers, stretches of DNA that help control when and where genes are expressed. Researchers found that depolarizing cultured neurons changed CBP occupancy at thousands of enhancers. A separate study in adult mice showed that neuronal activation changed chromatin accessibility, with many newly accessible regions associated with the AP-1 family of transcription factors. This helps explain why one electrical event can influence a coordinated set of genes and why a neuron and a muscle fiber can interpret a similar stimulus differently. Each cell starts with a different set of accessible regulatory regions and proteins available to act on them. [5,6] (pmc.ncbi.nlm.nih.gov, pubmed.ncbi.nlm.nih.gov)
3. Electrical activity can change DNA methylation
DNA methylation adds another layer of control. Methyl groups at particular DNA sites can influence how a gene is regulated, with effects that depend on the location of the mark and the proteins that recognize it.
In adult mice, electroconvulsive stimulation produced both gains and losses of DNA methylation at specific sites in the hippocampus. In related work, neuronal activity induced Gadd45b, which was required for activity-associated demethylation at regulatory regions of BDNF and FGF1 in that model. BDNF is relevant to Lionheart’s Brain Band™ research because it provides a concrete example of how neuronal activity can reach the regulatory DNA of a gene involved in plasticity. The animal studies used strong, synchronized neuronal activation. A particular noninvasive waveform would need its own measurements to establish whether it engages the same pathway. [7,8] (pmc.ncbi.nlm.nih.gov, pubmed.ncbi.nlm.nih.gov, lionheartlongevity.com)
4. Muscle contraction creates another route to the genome
When stimulation recruits muscle, the response includes electrical activation, calcium release, mechanical work, and a change in energy demand. In adult skeletal muscle fibers, researchers showed that electrical stimulation could drive HDAC4 out of the nucleus. HDAC4 can restrain the transcription factor MEF2, so its movement provides a route by which muscle activity can change transcription. In human muscle, an acute exercise study found changes in promoter methylation and expression of metabolic genes including PPARGC1A, which encodes PGC-1α, and PDK4. A separate resistance-training study found that some methylation changes persisted through a period without training. [9–11] (pubmed.ncbi.nlm.nih.gov, pubmed.ncbi.nlm.nih.gov, pubmed.ncbi.nlm.nih.gov)
Metabolism can contribute directly to chromatin regulation. Acetyl-CoA, for example, supplies the acetyl groups used to modify histones. Researchers have shown that ATP-citrate lyase can link nutrient metabolism to histone acetylation in mammalian cells. For a program such as BodStim™, this raises a testable question. If two electrical patterns produce the same amount of muscle work, do they still generate different calcium responses, metabolic changes, or chromatin marks? Matching the work helps distinguish the contribution of pulse timing from the contribution of contraction. [12] (pubmed.ncbi.nlm.nih.gov)
Klotho as a test case for bioelectric gene regulation
Alpha-Klotho is best known for its role as a co-receptor in FGF23 signaling, which helps regulate phosphate and mineral metabolism. The protein exists in membrane-associated and soluble forms. The original mouse studies brought Klotho to the attention of aging researchers because disruption of the KL gene produced a syndrome with several features resembling accelerated aging. Later work found that methylation of the KL promoter helps explain why the gene is expressed much more strongly in some tissues than in others. That makes Klotho particularly interesting for bioelectric research. Its expression is already known to depend, in part, on the regulatory state of the gene. [13–15] (pubmed.ncbi.nlm.nih.gov, pubmed.ncbi.nlm.nih.gov, pubmed.ncbi.nlm.nih.gov)
Leonhardt Ventures’ issued U.S. patent US 12,226,639 B2, Klotho Modulation, describes programmed electrical stimulation intended to increase Klotho expression or release. Lionheart also identifies licensed Klotho-expressing stem-cell technology as a separate part of its portfolio. The key experimental question is what a rise in soluble Klotho represents. It could reflect increased transcription of KL, greater release of protein already present, or a contribution from a different tissue. A study that measures circulating Klotho alongside tissue-specific KL RNA, membrane-associated protein, and regulatory DNA before and after stimulation could begin to distinguish those possibilities. Repeat measurements after stimulation stops would show whether any change in regulation persists. [18,21] (patents.google.com, investors.lionheartlongevity.
How Lionheart’s other protein targets fit
SIRT6, sirtuins, and sestrins. SIRT6 is one of the most direct links between the Lionheart portfolio and epigenetics because it can modify histones. Independent research showed that SIRT6 deacetylates histone H3 at lysine 9 in telomeric chromatin. Leonhardt Ventures’ issued U.S. patent US 12,752,107 B1, Modulation of SIRT, includes a claim directed to a defined electrical stimulation method for skeletal muscle that increases SIRT6 expression in the context of muscle atrophy. A mechanistic study could ask whether stimulation changes SIRT6 protein or activity and whether those changes alter its histone targets. Lionheart’s broader target guide also identifies SIRT1, SIRT3, and sestrins such as SESN2, which add related questions about mitochondrial metabolism and stress responses. [16,19,22] (pubmed.ncbi.nlm.nih.gov, patentsgazette.uspto.gov, investors.lionheartlongevity.
BDNF, Menin, follistatin, and IGF-1. These targets illustrate several levels of regulation. BDNF is an output of activity-dependent gene regulation and can influence neuronal plasticity. Menin participates in transcriptional regulation and is the subject of an announced Leonhardt Ventures provisional application focused on bioelectric enhancement in the brain. Follistatin binds activins and can modulate myostatin signaling, while IGF-1 participates in growth and survival signaling. In muscle research, measuring these proteins alongside force, fatigue, and recovery would help connect changes in expression to tissue function. [8,18,22,25] (pubmed.ncbi.nlm.nih.gov, patents.google.com, investors.lionheartlongevity.
SDF-1, PDGF, VEGF, eNOS, and HMGB1. Repair requires communication between cells as well as changes inside them. SDF-1, also called CXCL12, can guide migration of cells that express its receptor CXCR4. PDGF helps regulate the behavior of cells that organize repairing tissue. VEGF and eNOS are relevant to vascular growth and nitric oxide signaling, respectively. HMGB1 has distinct roles inside and outside the cell. Inside the nucleus it binds chromatin. Outside the cell it can serve as a damage signal whose effects depend on its form and the tissue context. Lionheart has announced a provisional application concerning bioelectric modulation of HMGB1. Research can ask whether an electrical intervention changes local protein release, cell recruitment, or both. [20,22,24] (patents.google.com, investors.lionheartlongevity.
COL17A1, tropoelastin, and Wnt signaling. In HairCell™ and related scalp research, COL17A1 is relevant to the environment that supports hair follicle stem cells. An independent study linked degradation of COL17A1 to age-related loss of those stem cells. Tropoelastin is the precursor to mature elastin and provides a separate target for studying tissue elasticity. Wnt signaling helps regulate stem-cell activity and tissue renewal. Lionheart’s HairCell materials describe a research strategy spanning these local tissue pathways and signals such as SDF-1, PDGF, VEGF, and eNOS. Lionheart and Leonhardt Ventures have also announced a provisional application for bioelectric hair repigmentation. Changes in hair density, pigmentation, and scalp elasticity are distinct endpoints and can be measured separately. [17,22,23] (pubmed.ncbi.nlm.nih.gov, investors.lionheartlongevity.
Thymulin, inflammatory signaling, and bone remodeling. Lionheart and its collaborators have announced work on thymulin, a thymic peptide involved in immune regulation. Separate orthodontic announcements illustrate why the desired direction of a signal can depend on the phase of treatment. RANKL promotes the formation and activity of bone-resorbing cells, while OPG limits RANKL signaling. BMP9 is relevant to bone-forming differentiation. Leonhardt Ventures has described proposed approaches involving these targets and, separately, IL-1β and PGE2 for orthodontic movement. A signal that helps initiate remodeling at one stage may need to give way to a different signal during stabilization. [22,26–28] (investors.lionheartlongevity.
What the patents cover and what experiments can establish
Lionheart reports commercialization and license rights to core Leonhardt Ventures technology, supplemented by third-party licenses and options. The public record includes issued patents such as Klotho Modulation, Modulation of SIRT, and US 11,052,247 B2, Skin Treatment System. The skin patent describes bioelectric stimulation in connection with several repair-related factors, including SDF-1, VEGF, PDGF, eNOS, HGF, follistatin, and tropoelastin. The scope of protection comes from the granted claims. Lionheart News also describes application-stage work on hair repigmentation, HMGB1, Menin, thymulin, polylaminin-related approaches, and bioelectrically enhanced exosomes. These announcements identify research and filing activity. Evidence for efficacy, mechanism, and durability comes from studies designed to measure those outcomes. [18–28] (investors.lionheartlongevity.
What would show that a waveform truly reprogrammed a tissue?
A convincing study would connect the electrical input to a change in a defined cell population, then follow that change after stimulation ends.
- Measure the signal the tissue receives. Specify electrode geometry, amplitude, pulse width, frequency, burst spacing, and session duration. Measure the relevant electrical, calcium, or contractile response in the target tissue.
- Find the cells that respond. Determine which cells increase or decrease RNA or protein production. For secreted proteins such as soluble Klotho, establish the source where possible.
- Measure the regulatory change. Pair RNA and protein measurements with assays for chromatin accessibility, histone marks, or DNA methylation. This can help distinguish fresh gene expression from protein release and changes within a cell from changes in which cells are present.
- Test the mechanism and follow it over time. Compare sham and active waveforms, match contraction where relevant, and test whether blocking a candidate pathway changes the response. Then measure whether the molecular change and its functional effect persist after stimulation stops.
The most informative result would connect a defined waveform to a measured cellular response, a change at a specific regulatory region, and a functional effect that depends on that change. That is how a promising observation such as increased Klotho, BDNF, or SIRT6 can become evidence for a reproducible form of bioelectric epigenetic reprogramming.
Scientific references
- Deisseroth K, Heist EK, Tsien RW. Translocation of calmodulin to the nucleus supports CREB phosphorylation in hippocampal neurons. Nature. 1998. Full text
- Bito H, Deisseroth K, Tsien RW. CREB phosphorylation and dephosphorylation as a calcium- and stimulus-duration-dependent switch for hippocampal gene expression. Cell. 1996. PubMed
- Dolmetsch RE, Xu K, Lewis RS. Calcium oscillations increase the efficiency and specificity of gene expression. Nature. 1998. Article
- Impey S, et al. Phosphorylation of CBP mediates transcriptional activation by neural activity and CaM kinase IV. Neuron. 2002. PubMed
- Kim TK, et al. Widespread transcription at neuronal activity-regulated enhancers. Nature. 2010. Full text
- Su Y, et al. Neuronal activity modifies the chromatin accessibility landscape in the adult brain. Nature Neuroscience. 2017. PubMed
- Guo JU, et al. Neuronal activity modifies the DNA methylation landscape in the adult brain. Nature Neuroscience. 2011. Full text
- Ma DK, et al. Neuronal activity-induced Gadd45b promotes epigenetic DNA demethylation and adult neurogenesis. Science. 2009. PubMed
- Liu Y, Randall WR, Schneider MF. Activity-dependent and -independent nuclear fluxes of HDAC4 mediated by different kinases in adult skeletal muscle. Journal of Cell Biology. 2005. PubMed
- Barrès R, et al. Acute exercise remodels promoter methylation in human skeletal muscle. Cell Metabolism. 2012. PubMed
- Seaborne RA, et al. Human skeletal muscle possesses an epigenetic memory of hypertrophy. Scientific Reports. 2018. PubMed
- Wellen KE, et al. ATP-citrate lyase links cellular metabolism to histone acetylation. Science. 2009. PubMed
- Kuro-o M, et al. Mutation of the mouse klotho gene leads to a syndrome resembling ageing. Nature. 1997. PubMed
- Kurosu H, et al. Regulation of fibroblast growth factor-23 signaling by klotho. Journal of Biological Chemistry. 2006. PubMed
- Azuma M, et al. Promoter methylation confers kidney-specific expression of the Klotho gene. FASEB Journal. 2012. PubMed
- Michishita E, et al. SIRT6 is a histone H3 lysine 9 deacetylase that modulates telomeric chromatin. Nature. 2008. PubMed
- Matsumura H, et al. Hair follicle aging is driven by transepidermal elimination of stem cells via COL17A1 proteolysis. Science. 2016. PubMed
Patent records and company disclosures
- Leonhardt Ventures LLC. Klotho Modulation. U.S. Patent 12,226,639 B2. Patent record
- Leonhardt Ventures LLC. Modulation of SIRT. U.S. Patent 12,752,107 B1. USPTO Official Gazette
- Skin Treatment System. U.S. Patent 11,052,247 B2. Patent record
- Lionheart Health. Intellectual Property Portfolio
- Lionheart Health. Protein & Signaling Targets
- Lionheart Health. HairCell™ technology and bioelectric hair repigmentation provisional filing announcement
- Lionheart Health. HMGB1 provisional application announcement
- Leonhardt Ventures. Menin provisional filing announcement
- Leonhardt Ventures, Lionheart Health, and CancerCell. Thymulin provisional application preparation announcement
- Leonhardt Ventures. RANKL, OPG, and BMP9 orthodontic platform announcement and IL-1β and PGE2 provisional application announcement
- Lionheart Health. Polylaminin-related provisional filing announcement and bioelectrically enhanced exosomes research and provisional filing announcement