Executive Summary
Lionheart Health proposes a translational research initiative to determine whether increasing alpha-Klotho can reduce seizure-associated brain injury, neuroinflammation and cognitive decline—and potentially lower seizure susceptibility or burden.
The current evidence suggests that epilepsy may create a state of localized Klotho deficiency, particularly within the hippocampus. Restoring Klotho could interrupt several processes that contribute to epileptogenesis and epilepsy progression, including chronic inflammation, oxidative stress, ferroptosis, neuronal apoptosis and impaired synaptic plasticity.
The evidence remains primarily observational and preclinical. Klotho should therefore be investigated initially as an adjunct to standard antiseizure therapy, not as a replacement for approved medications.
Scientific Relationship Between Klotho and Epilepsy
1. Klotho is reduced in human temporal-lobe epilepsy
Resected hippocampal tissue from patients with temporal-lobe epilepsy and hippocampal sclerosis showed:
- Significantly reduced KL gene expression
- Markedly increased TNF
- Increased NF-κB
- An inverse relationship between hippocampal Klotho and the inflammatory TNF/NF-κB pathway
These findings support a model in which persistent seizures and neuroinflammation suppress Klotho, while reduced Klotho may leave neurons increasingly vulnerable to inflammation, synaptic loss and degeneration.
2. Higher circulating Klotho is associated with lower epilepsy prevalence
A 2025 analysis of 36,578 adults in the U.S. NHANES database found that higher serum Klotho was associated with lower epilepsy prevalence. The reported adjusted odds ratio was 0.74, although the relationship was nonlinear and appeared stronger in men and adults younger than 65.
This was a cross-sectional association and cannot establish that low Klotho causes epilepsy or that raising Klotho prevents seizures. It does, however, provide human population-level support for further prospective research.
3. Klotho suppresses epilepsy-associated neuroinflammation
In a lithium-pilocarpine rat model of temporal-lobe epilepsy, hippocampal Klotho declined after epilepsy induction. AAV-mediated Klotho overexpression:
- Reduced hippocampal neuronal injury
- Improved cognitive performance
- Decreased NLRP3, caspase-1 and IL-1β
- Activated the antioxidant regulator Nrf2
Blocking Nrf2 substantially reversed Klotho’s benefits, supporting a Klotho–Nrf2–NLRP3 mechanism. The study primarily supports protection against epilepsy-associated inflammation and cognitive damage rather than proving direct seizure elimination.
4. Klotho protects against ferroptosis and oxidative injury
A separate temporal-lobe epilepsy rat study found that hippocampal Klotho overexpression:
- Reduced iron accumulation
- Increased glutathione and GPX4
- Lowered reactive oxygen species
- Regulated ferroportin and divalent metal transporter 1
- Reduced ferroptotic neuronal injury
- Improved epilepsy-associated cognitive deficits
This positions Klotho as a potential protector against the progressive neuronal damage that accompanies recurrent seizures.
5. Pharmacologic Klotho activation may reduce seizures
A 2025 study evaluated a modified physostigmine analogue in PTZ-kindled mice. The compound increased Klotho expression and produced dose-related anticonvulsant, neuroprotective and cognitive benefits while reducing inflammatory and apoptotic markers.
Important limitations remain: the investigators did not conclusively prove that Klotho activation was solely responsible for seizure suppression, and long-term safety, pharmacokinetics and performance across multiple epilepsy models remain unknown.
Proposed Therapeutic Hypothesis
Lionheart’s central hypothesis is:
Restoring Klotho signaling may increase neural resilience and modify the inflammatory, oxidative and degenerative environment that promotes recurrent seizures, hippocampal injury and epilepsy-associated cognitive decline.
A Klotho therapy could potentially address five interconnected targets:
- Neuroinflammation: Suppression of TNF/NF-κB and NLRP3/IL-1β signaling.
- Oxidative stress: Activation of Nrf2 and endogenous antioxidant defenses.
- Ferroptosis: Preservation of GPX4, glutathione and normal iron handling.
- Neuronal survival: Reduction of caspase-3, BAX-mediated apoptosis and excitotoxic injury.
- Cognitive resilience: Preservation of hippocampal synaptic plasticity, learning and memory.
Systemic recombinant Klotho has enhanced cognition in aged nonhuman primates, demonstrating that peripherally administered Klotho can influence brain function even though the intact protein does not appear to cross the blood–brain barrier. The primate study also found benefits at a lower dose but not at higher doses, emphasizing the importance of careful dose optimization.
Lionheart Translational Research Program
Phase 1 — Human Biomarker Study
Enroll patients with controlled epilepsy, drug-resistant epilepsy and matched controls.
Measure:
- Serum and, where clinically appropriate, cerebrospinal-fluid Klotho
- Seizure frequency and severity
- EEG abnormalities and epileptiform-discharge burden
- Cognitive performance, memory, mood and sleep
- TNF-α, IL-1β, IL-6, NLRP3-related markers
- Oxidative stress, glutathione, GPX4 and iron-metabolism markers
- Renal function, calcium, phosphate and other safety laboratories
Primary objective: determine whether Klotho levels correlate with epilepsy subtype, seizure burden, cognitive impairment or inflammatory status.
Phase 2 — Preclinical Intervention Study
Test Lionheart-selected Klotho-elevating strategies in at least two epilepsy models, such as PTZ kindling and lithium-pilocarpine temporal-lobe epilepsy.
Potential research arms could include:
- Recombinant soluble alpha-Klotho
- Klotho-enhancing small molecules
- Targeted Klotho gene or mRNA expression
- Lionheart bioelectric Klotho-expression protocols
- Combination treatment with standard antiseizure medication
Required outcomes should include continuous video-EEG, spontaneous seizure frequency, seizure duration, mortality, cognition, hippocampal pathology and mechanistic biomarkers.
Phase 3 — Early Clinical Feasibility Trial
Following adequate safety and preclinical efficacy, conduct a randomized, sham- or placebo-controlled adjunctive study in adults with persistent focal seizures.
Potential primary endpoint:
- Percentage reduction in monthly seizure frequency
Important secondary endpoints:
- Responder rate of at least 50%
- Interictal EEG burden
- Memory and executive function
- Mood, fatigue and sleep
- Serum Klotho and inflammatory biomarkers
- Safety and interaction with antiseizure medications
Strategic Conclusion
Klotho is not yet a proven antiseizure therapy. Nevertheless, converging human-tissue, epidemiological and animal evidence supports a scientifically credible relationship between Klotho deficiency, neuroinflammation, oxidative injury, ferroptosis and epilepsy-associated cognitive decline.
The strongest near-term opportunity for Lionheart is to develop Klotho as a neuroprotective and disease-modifying adjunct that may:
- Protect hippocampal neurons from repeated seizure injury
- Preserve cognition and memory
- Reduce inflammatory and oxidative drivers of epileptogenesis
- Potentially reduce seizure severity or frequency when combined with standard treatment
Principal Supporting Papers
- Teocchi et al. “Hippocampal gene expression dysregulation of Klotho, nuclear factor kappa B and tumor necrosis factor in temporal lobe epilepsy patients.” Journal of Neuroinflammation, 2013.
- Xiang et al. “Klotho ameliorated cognitive deficits in a temporal lobe epilepsy rat model by inhibiting ferroptosis.” Brain Research, 2021.
- Xiang et al. “Klotho alleviates NLRP3 inflammasome-mediated neuroinflammation in a temporal lobe epilepsy rat model by activating the Nrf2 signaling pathway.” Epilepsy & Behavior, 2022.
- Chen et al. “Serum Klotho levels and epilepsy among U.S. adults in the NHANES 2013–2016.” Scientific Reports, 2025.
- Dahalia et al. “In vitro and in-vivo exploration of physostigmine analogues to understand the mechanistic crosstalk between Klotho and targets for epilepsy.” Frontiers in Pharmacology, 2025.
- Castner et al. “Longevity factor Klotho enhances cognition in aged nonhuman primates.” Nature Aging, 2023.
Lionheart Health Multi-Arm Klotho Elevation Research Initiative
Research Objective
Lionheart Health proposes to evaluate a portfolio of complementary technologies designed either to:
- Stimulate the body’s endogenous production of alpha-Klotho;
- Deliver soluble or recombinant Klotho;
- Establish cells or tissues that continuously express Klotho; or
- Support Klotho expression through exercise, nutrition and metabolic optimization.
For epilepsy research, the central hypothesis is that restoring Klotho may improve neuronal resilience, reduce neuroinflammation and oxidative injury, protect hippocampal function and potentially reduce the biological processes that promote seizure recurrence.
Klotho is not presently an approved epilepsy treatment. Each modality should be classified according to its scientific maturity and regulatory status.
Proposed Research Arms
Arm 1 — Klotho Patch™
The Klotho Patch™ would be developed as a repeated-use, minimally invasive delivery or stimulation platform.
Potential configurations include:
- Bioelectric patch intended to stimulate endogenous Klotho expression;
- Microneedle patch containing recombinant Klotho, a Klotho-derived peptide, mRNA or another Klotho-promoting agent;
- Hydrogel-forming microneedle patch providing sustained release;
- Patch combined with iontophoresis or another electrically assisted delivery method;
- Patch containing nutraceutical ingredients intended to support endogenous Klotho production.
A conventional passive adhesive patch is unlikely to deliver meaningful quantities of intact Klotho protein because Klotho is a large biological molecule and the skin is an effective barrier. Microneedles, iontophoresis or another active delivery system would therefore be more scientifically credible for direct protein or nucleic-acid delivery. Microneedles have been studied for transdermal delivery of proteins, peptides, vaccines and nucleic acids.
Development status: Prototype and preclinical delivery validation.
Arm 2 — Klotho Pod I™
Klotho Pod I™ may serve as Lionheart’s localized or wearable Klotho-elevation platform.
The research configuration could combine:
- Local bioelectric stimulation;
- A Klotho Patch or topical delivery interface;
- Local photobiomodulation or PEMF;
- Temperature, oxygenation or circulation support;
- Biomarker-guided adjustment of treatment intensity;
- Targeting of Klotho-producing or Klotho-responsive tissues.
This arm should determine whether a compact, repeatable system can produce measurable changes in circulating Klotho without requiring systemic biological therapy.
Development status: Investigational wellness and biomarker research platform.
Arm 3 — Klotho Pod II™ Rejuvenation Chamber
Klotho Pod II™ is envisioned as Lionheart’s noninvasive, multimodality whole-body Klotho optimization chamber.
The integrated protocol may include:
- BodStim™ whole-body neuromuscular stimulation;
- Frequency-specific bioelectric signaling;
- Photobiomodulation;
- PEMF;
- Circulation and muscle activation;
- Relaxation and autonomic-balancing modalities;
- Brain Band™ and Second Brain™ components;
- Klotho Patch™ or other noninvasive delivery options;
- Real-time physiological and biomarker monitoring.
The purpose is to test whether multiple mild interventions acting together produce a larger and more durable Klotho response than any one modality alone.
Development status: Investigational multimodality research system; individual components must be evaluated separately before attributing any combined effect specifically to Klotho.
Arm 4 — BodStim™ Klotho Stimulation
BodStim™ uses whole-body EMS or neuromuscular electrical stimulation to reproduce and augment muscle-contraction signals associated with exercise.
Exercise is currently one of the most scientifically supportable non-drug methods for increasing soluble Klotho. A randomized 12-week exercise trial in middle-aged adults reported increases in circulating soluble Klotho, and aerobic exercise has also been associated with increased plasma Klotho and improved arterial compliance. A systematic review concluded that exercise generally increases Klotho across studied populations, although responses vary by age, health status and exercise protocol.
Lionheart should test whether its specific BodStim waveforms provide benefits beyond those produced by comparable voluntary exercise. The BodStim platform is described by Lionheart as a whole-body EMS system incorporating bioelectric programs intended to support Klotho and other muscle-related protein expression.
Recommended study groups:
- Voluntary exercise alone;
- Standard EMS;
- BodStim proprietary Klotho program;
- BodStim plus exercise;
- Sham stimulation.
Development status: Suitable for early human biomarker studies, provided claims remain limited to measured study outcomes.
Arm 5 — Brain Band™ Targeted Klotho Stimulation
Brain Band™ may be evaluated as a targeted neuromodulation platform intended to influence:
- Choroid-plexus Klotho production;
- Hippocampal resilience;
- Nrf2 antioxidant signaling;
- Neuroplasticity;
- Neuroinflammation;
- Sleep and autonomic regulation;
- Cognitive recovery following seizures.
Klotho has demonstrated cognition-enhancing and neuroprotective effects in animal models, including protection against glutamate toxicity and enhancement of synaptic function. Systemic Klotho administration also improved cognition in aged nonhuman primates at a lower tested dose, although the higher dose did not produce the same benefit.
However, there is not yet sufficient evidence that transcranial stimulation directly increases brain or circulating Klotho in humans. That relationship must be demonstrated experimentally rather than assumed.
Epilepsy Safety Requirement
Any Brain Band protocol for epilepsy must be developed with an epileptologist, continuous or repeated EEG monitoring and conservative stopping rules. Photic or flashing-light stimulation requires particular caution because visually provocative stimuli can trigger seizures in a subset of people with epilepsy.
Development status: Exploratory, IRB-supervised neuromodulation research.
Arm 6 — Second Brain™ Gut–Brain Klotho Stimulation
Second Brain™ may target the gut–brain, immune and autonomic pathways that indirectly influence Klotho expression and seizure susceptibility.
Potential mechanisms to study include:
- Vagus-nerve and autonomic modulation;
- Reduction of systemic inflammation;
- Improvement in gut-barrier integrity;
- Changes in microbiome composition;
- Improved insulin sensitivity;
- Reduced inflammatory cytokine signaling;
- Kidney–gut–brain communication;
- Increased endogenous Klotho expression secondary to improved metabolic and inflammatory health.
Vagus-nerve stimulation is already an established adjunctive treatment for some patients with drug-resistant epilepsy, but its clinical benefits have not been proven to result from Klotho elevation.
Lionheart should therefore measure both seizure outcomes and Klotho biomarkers to determine whether Second Brain produces a Klotho-mediated effect, an independent neuromodulatory effect or both.
Development status: Mechanistic human feasibility research.
Arm 7 — Recombinant Klotho Protein Infusions or Injections
Direct administration of soluble recombinant alpha-Klotho could provide a rapid but temporary increase in circulating Klotho.
Potential delivery routes include:
- Intravenous infusion;
- Subcutaneous injection;
- Intramuscular injection;
- Intrathecal or intracerebroventricular delivery in preclinical research only;
- Sustained-release hydrogel;
- Nanoparticle or microneedle delivery.
A single low-dose systemic administration improved memory in aged nonhuman primates, supporting the concept that peripheral Klotho treatment can influence brain function. The dose response was not linear, emphasizing the need for careful pharmacokinetic and dose-ranging work.
Research priorities include:
- Half-life and tissue distribution;
- Dose–response relationship;
- Immune reactions and antibody formation;
- Calcium, phosphate, FGF23 and vitamin-D regulation;
- Effects on seizure threshold;
- Combination with antiseizure medication.
Development status: Preclinical development followed by an IND-regulated clinical program.
Arm 8 — Klotho-Expressing Stem-Cell Infusions or Injections
Mesenchymal stromal cells or other appropriate cell populations could be genetically engineered to express secreted Klotho and function as temporary biological Klotho-production centers.
Possible administration routes include:
- Intravenous infusion;
- Intramuscular injection;
- Local injection near an injured tissue;
- Intrathecal administration in advanced preclinical studies;
- Implantation within a protective nutrient hydrogel;
- Delivery with local bioelectric stimulation intended to improve cell survival and controlled protein release.
Preclinical studies have shown that Klotho-modified mesenchymal cells can increase Klotho delivery and improve outcomes in models of kidney injury, pulmonary hypertension and neuroinflammatory disease. These findings support continued research but do not establish safety or effectiveness in epilepsy or humans.
Key safety questions include:
- Cell identity, purity and genetic stability;
- Tumorigenicity;
- Uncontrolled or excessive Klotho expression;
- Immune rejection;
- Thrombosis and embolic risk;
- Infection;
- Biodistribution;
- Duration and reversibility of expression.
Development status: Advanced preclinical and IND-regulated cell-therapy research.
Arm 9 — Klotho-Expressing Cells in Nutrient Hydrogel
Rather than freely infusing cells, Lionheart may encapsulate Klotho-expressing cells in a biocompatible nutrient hydrogel.
Potential advantages include:
- Sustained local Klotho secretion;
- Improved cell viability;
- Reduced migration of cells to unintended organs;
- Retrievability of the cell construct;
- Localized treatment near the kidney, brain-supporting tissues or another target;
- Controlled activation with bioelectric stimulation;
- Reduced frequency of repeat infusions.
This approach could ultimately be integrated into a Klotho Implant™, refillable pump or Lionheart multi-organ regeneration system.
Development status: Preclinical biomaterials, cell-survival and controlled-release research.
Arm 10 — Klotho Gene Therapy
Klotho gene therapy could provide longer-duration expression than protein injection or stimulation alone.
Potential platforms include:
- AAV carrying secreted Klotho;
- AAV carrying full-length Klotho;
- Nonviral plasmid DNA;
- Lipid-nanoparticle mRNA;
- Tissue-targeted polymeric nanoparticles;
- Ex vivo modification of autologous cells;
- Regulatable or externally activated gene-expression systems.
AAV-mediated Klotho expression has produced multi-organ benefits in aging and disease models. Other animal studies have reported improvements in vascular, renal, metabolic and aging-related outcomes following Klotho gene delivery.
For epilepsy, an initial program should favor controlled or tissue-targeted expression rather than permanent, unregulated whole-body overexpression.
Critical development requirements include:
- Vector biodistribution;
- Dose control;
- Immune response to the vector;
- Liver and kidney safety;
- Calcium–phosphate monitoring;
- Durability of expression;
- Reversibility or shutoff capability;
- Reproductive and germline-exposure precautions;
- Continuous seizure and EEG monitoring.
Development status: Preclinical gene-therapy program requiring FDA authorization before U.S. human administration.
Arm 11 — Bioelectric-Enhanced Klotho Nanoflowers
Lionheart’s Klotho Nanoflower platform may combine flower-shaped nanoparticles, Klotho-related biological payloads and frequency-specific bioelectric activation.
Potential payloads include:
- Recombinant Klotho protein;
- Klotho-derived peptides;
- KL plasmid DNA;
- Klotho mRNA;
- CRISPR activation components;
- Antioxidant or mitochondrial-support compounds;
- Stem-cell-homing signals.
Lionheart describes the investigational platform as using bioelectric fields to promote nanoflower uptake, intracellular transport and activation of biological payloads.
Independent preclinical research has demonstrated that nanoparticle-mediated Klotho gene delivery can protect renal tissue and modify disease-associated signaling, supporting the broader feasibility of nanoparticle Klotho delivery.
For epilepsy, candidate targeting strategies could include the hippocampus, choroid plexus, cerebral vasculature or peripheral organs that regulate systemic inflammation.
Development status: Early preclinical nanomedicine and combination-product research.
Arm 12 — Klotho-Supporting Supplements
The supplement arm should be positioned as supporting the body’s endogenous Klotho expression rather than supplying intact oral Klotho protein.
Potential categories for controlled evaluation include:
- Vitamin D correction when deficiency is documented;
- Alpha-ketoglutarate formulations;
- Polyphenol-rich botanical combinations;
- Antioxidants;
- Mitochondrial-support ingredients;
- Anti-inflammatory nutrients;
- Kidney-supportive micronutrients;
- Lionheart-selected KlothaMax™ or comparable formulations;
- Rejuvant or other AKG-based protocols;
- Personalized supplementation based on laboratory deficiencies.
A clinical study of vitamin-D supplementation has evaluated effects on alpha-Klotho and FGF23, but supplement effects on Klotho are not consistent enough to justify broad claims without product-specific testing.
Every formulation should be independently tested against placebo using the same validated Klotho assay. Supplements must also be reviewed for interactions with antiseizure medications and for ingredients that could lower seizure threshold.
Development status: Human wellness and biomarker research, with product-specific substantiation required.
Arm 13 — Klotho-Promoting Diet
Lionheart may employ a personalized Mediterranean-style, anti-inflammatory and metabolically supportive diet as a foundational Klotho arm.
The diet may emphasize:
- Vegetables and leafy greens;
- Berries and lower-glycemic fruits;
- Legumes;
- Nuts and seeds;
- Extra-virgin olive oil;
- Fish and appropriate omega-3 sources;
- Adequate but not excessive protein;
- Fiber and microbiome-supportive foods;
- Reduced refined carbohydrates;
- Reduced ultra-processed foods;
- Weight optimization;
- Avoidance of excessive phosphate additives;
- Adequate hydration and kidney support.
Greater Mediterranean-diet adherence has been associated with higher soluble Klotho levels in observational analyses. Diet quality, body composition and lean-mass measures may all influence Klotho, but these studies do not prove that diet alone directly causes a therapeutic Klotho increase.
Phosphate restriction should not be applied indiscriminately. It should be guided by renal function, serum phosphate, calcium, vitamin D, parathyroid hormone and nutritional status.
Development status: Core human research and standard-of-care lifestyle optimization arm.
Arm 14 — Integrated Combination Protocol
The final Lionheart research arm may evaluate a complete multimodality Klotho-Up™ protocol:
- Klotho-promoting diet;
- Personalized supplements;
- BodStim two to three times weekly;
- Klotho Pod II sessions;
- Klotho Patch;
- Brain Band or Second Brain when appropriate;
- Sleep, metabolic and inflammation optimization;
- Recombinant Klotho or peptides in an advanced investigational arm;
- Klotho-expressing cells or gene therapy only under the applicable regulatory authorization.
The combination arm should only begin after the contribution and safety of the principal components have been characterized. Otherwise, it will be impossible to identify which modality produced the observed benefit or adverse event.
Recommended Development Tiers
Tier 1 — Lower-Risk Human Biomarker Arms
- Diet;
- Supplements;
- Voluntary exercise;
- BodStim;
- Klotho Pod I;
- Klotho Pod II;
- Non-delivery Klotho Patch;
- Sleep and metabolic optimization.
Tier 2 — IRB-Supervised Experimental Arms
- Microneedle Klotho Patch;
- Brain Band targeted stimulation;
- Second Brain gut–brain stimulation;
- Recombinant Klotho protein;
- Klotho peptides;
- Hydrogel delivery;
- Combination neuromodulation protocols.
Tier 3 — IND-Regulated Biological Arms
- Klotho-expressing stem cells;
- Genetically modified cell implants;
- AAV or other Klotho gene therapy;
- mRNA or DNA nanoparticles;
- Klotho nanoflowers carrying gene or biological payloads.
FDA states that regenerative cell products and gene- or tissue-based products intended to treat disease generally require regulatory authorization and FDA-supervised clinical development before they may be marketed in the United States.
Common Measurements Across All Arms
To permit valid comparisons, every arm should use the same core measurements:
- Serum soluble alpha-Klotho;
- Repeat Klotho measurement using a validated assay;
- FGF23, phosphate, calcium and vitamin D;
- Renal and hepatic function;
- Seizure frequency and duration;
- Continuous, ambulatory or scheduled EEG;
- Interictal epileptiform-discharge burden;
- Antiseizure-medication levels;
- Memory and executive-function testing;
- Depression, anxiety, fatigue and sleep;
- TNF-α, IL-1β, IL-6 and CRP;
- Nrf2-related antioxidant markers;
- Glutathione, GPX4 and iron-metabolism markers;
- Adverse events and serious adverse events.
Epilepsy-Specific Safety Principles
- Klotho interventions must remain adjunctive to established epilepsy care.
- Participants should not discontinue or reduce antiseizure medications without their treating neurologist.
- Brain stimulation must use epilepsy-specific safety parameters and EEG oversight.
- Participants should be screened for photosensitivity before any flashing-light protocol.
- Cell, gene, recombinant-protein and nanoparticle arms require formal toxicology and regulatory review.
- Calcium, phosphate, FGF23, vitamin D and kidney function must be monitored because Klotho participates in mineral metabolism.
- Each protocol should include predetermined seizure-related stopping rules.
Strategic Positioning
Lionheart Health’s principal advantage is the ability to compare several fundamentally different approaches within one coordinated development platform:
Lifestyle activation + bioelectric endogenous expression + targeted neuromodulation + direct protein delivery + cellular production + gene-based expression + nanotechnology delivery.
This creates an opportunity to determine not only whether Klotho can be elevated, but which method produces the safest, most durable and most clinically meaningful response for epilepsy, cognition and broader healthspan.