The Innovative Project Award provides $200,000 over two years and is deliberately designed to back high-risk, high-reward science. Eligible projects spanned the full breadth of biomedical inquiry, from basic laboratory studies of cells and molecules to clinical investigations and population health research, and applicants could propose work across the entire human lifespan, from children and adolescents through middle age and late life. The prioritization of innovation over incremental safety reflects a conscious strategy by both societies: rather than extending established research programs, they wanted to seed ideas that conventional funding mechanisms might consider too speculative, but that could open entirely new avenues for preventing stroke and heart disease in people living with chronic headache disorders.

This year’s co-funded award goes to Juliana Navia Pelaez, Ph.D., an assistant professor at St. Louis University School of Medicine in St. Louis, Missouri, for a project titled “Neurogenic Priming of Perivascular Macrophages by Migraine-Associated Peptides Increases Stroke Risk.” Her work targets a question that has long puzzled clinicians: why some people who suffer from migraine face a higher risk of stroke. Researchers have traditionally concentrated on the blood vessels themselves, since during a migraine attack these vessels may tighten, swell, or alter the way blood moves through them. But attention is now shifting toward another, less obvious player — the brain’s own resident immune cells, which stand guard along the vessels and help protect neural tissue from injury.

Pelaez’s central hypothesis is that repeated migraine attacks may push these vascular immune cells into a state of chronic overactivation. “If migraines happen over and over, these immune cells might stay ‘on’ for too long. When this happens, they might accidentally make the blood vessels weaker,” Pelaez said. “If the vessels get weaker, the brain might have a harder time protecting itself from a stroke.” In practical terms, the project will map which signals released during a migraine activate these brain immune cells, and then compare how the cells respond to a subsequent stroke in animals that have experienced migraine-like events versus those that have not. The comparison is designed to isolate the specific contribution of migraine biology to stroke vulnerability, rather than attributing everything to shared risk factors such as hypertension or smoking.

The translational implications could be considerable. “If our idea is right,” Pelaez added, “this could help scientists find new ways to prevent strokes in people with chronic migraine, maybe by calming these overactive brain cells or blocking the signals that bother them.” Such a strategy would represent a fundamentally different approach to stroke prevention in this population. Instead of managing vascular risk factors after the fact, clinicians might one day intervene directly on the neuroimmune signaling cascade that links repeated migraine attacks to cumulative vessel damage — a possibility that could reshape how neurologists and cardiologists jointly manage patients with frequent or chronic migraine.

The Heart Association’s three additional awards extend the same investigative ambition across distinct mechanistic frontiers. The first goes to Andrea M. Harriott, M.D., Ph.D., an assistant professor in neurology at Massachusetts General Hospital in Boston, for a study titled “The Impact Of Cortical Spreading Depolarizations On Endothelial Phenotype, Neovascularization, and Collateral Remodeling.” Migraine with aura has long been epidemiologically linked to stroke and heart attack, and earlier thinking held that this association might simply reflect an elevated burden of conventional risk factors among people with aura. Harriott’s project will test a more direct possibility: that the long-term exposure to the mechanisms underlying migraine aura — cortical spreading depolarizations, waves of altered electrical and metabolic activity that sweep across the brain — causes lasting structural and functional harm to cerebral blood vessels. Her team will examine protein-level changes in the cells lining vessel walls, the formation of new blood vessels, and the remodeling of collateral circulation that the brain relies on when its primary supply routes are compromised.

The second award supports Matthew T. Bender, M.D., an associate professor at the University of Rochester in Rochester, New York, whose project, “Defining The Glymphatic-Hemodynamic Axis In IIH: A Novel Multi-Scale Quantitative MRI Framework,” addresses idiopathic intracranial hypertension, or IIH. This headache disorder disproportionately affects overweight women of childbearing age, and clinicians believe it arises from elevated pressure inside the skull driven by excess cerebrospinal fluid. Recent evidence, however, suggests the disorder may also involve dysfunction of the brain’s waste clearance network, known as the glymphatic system. Bender’s team will develop a new magnetic resonance imaging framework capable of visualizing both the glymphatic system and cerebral blood flow without contrast dye or invasive procedures. By scanning IIH patients before and after venous stenting — a procedure that opens narrowed veins draining the brain — the researchers hope to determine how restoring venous outflow affects waste clearance and hemodynamics, potentially establishing imaging markers that could guide treatment decisions.

The third award funds Neil Dani, Ph.D., an assistant professor at Vanderbilt University in Nashville, Tennessee, for “Illuminating Nociceptive And Analgesic Mechanisms Of Choroid Plexus In Migraine And Obesity Models.” Dani’s work responds to the rising prevalence of obesity-related migraine, a clinically important overlap given that both conditions share inflammatory signaling pathways. His focus is the choroid plexus, a small structure inside the brain’s ventricles that produces cerebrospinal fluid and acts as a highly regulated interface, or gate, between the body and the central nervous system. The working hypothesis is that inflammatory signals generated in the body during obesity may travel through the cerebrospinal fluid via the choroid plexus, sensitizing pain circuits and increasing migraine frequency or severity. The project will characterize inflammation patterns in this structure, identify cellular rewiring and immune signaling changes associated with obesity and migraine, and test whether clinically approved drugs — including acetazolamide, a long-standing medication already used in related contexts — can reduce both swelling and pain signaling in experimental models.

Taken together, the four projects sketch a research landscape in which headache disorders and vascular medicine are no longer treated as separate silos. Each award interrogates a different node of the heart-brain axis: immune cells that patrol cerebral vessels, the electrical storms of migraine aura and their vascular aftermath, the interplay of intracranial pressure and brain waste clearance, and the inflammatory crosstalk between metabolism, body-wide immune signaling, and pain. The funders emphasize that these efforts demonstrate the shared commitment of the American Headache Society and the American Heart Association to advancing brain health research, and the co-funding structure itself may prove as influential as the science it supports, encouraging cross-disciplinary training, shared patient cohorts, and unified endpoints across neurology and cardiology.

The initiative also sits within a far larger funding tradition. Supporting scientific research and discovery through programs of this kind is a cornerstone of the century-old American Heart Association’s lifesaving mission. Since 1949, the Association has invested more than $6.3 billion in cardiovascular, cerebrovascular, and brain health research, making it the single largest non-profit, non-governmental supporter of heart and brain health research in the United States. Officials note that knowledge generated through this funding continues to save lives and directly affect millions of people across the country and around the world. For the millions who live with migraine and other headache disorders — and for the clinicians who care for them — the new awards offer something more immediate than long-term promise: a concrete, funded research agenda that treats the pounding in the head and the health of the heart and brain’s vessels as inseparable parts of the same biological story.

Subject of Research: Funded research projects investigating the links between migraine, headache disorders, and cardiovascular and cerebrovascular health

Article Title: New scientific research projects study heart/brain connection to migraines, headaches

Article References: New scientific research projects study heart/brain connection to migraines, headaches. (n.d.). Original publication

Image Credits: AI Generated

DOI: Not provided

Keywords: migraine, stroke, American Heart Association, American Headache Society, cerebrovascular health, cortical spreading depolarization, idiopathic intracranial hypertension, glymphatic system, choroid plexus, perivascular macrophages, obesity-related migraine, research funding

The challenge that the method, called VINE-seq, was designed to solve is fundamentally mechanical. Most single-nucleus sequencing workflows begin by gently breaking brain tissue apart and releasing nuclei from individual cells. That approach works reasonably well for neurons and glia, but vascular cells are a different story. They are encased within a tough basement membrane, a dense meshwork of proteins that resists standard dissociation methods. As a result, when researchers homogenize brain tissue for single-nucleus profiling, vascular cells are systematically lost, depleted, or damaged. The consequence has been a major blind spot in the single-cell atlases of the human brain that now underpin much of modern neuroscience: the very cells that form the blood-brain barrier and regulate cerebral blood flow are dramatically underrepresented in the datasets used to study health and disease.

VINE-seq, short for vessel isolation and nucleus extraction for sequencing, tackles the problem head-on by isolating the vessels themselves before any attempt is made to extract their genetic material. The workflow begins with fresh or frozen brain tissue, either human or mouse, which is homogenized under conditions that preserve vascular integrity. The homogenate is then subjected to dextran-based density-gradient centrifugation, a technique that exploits differences in buoyant density to separate intact vessels from the surrounding myelin and the parenchymal fraction containing neurons and glia. The vessels recovered by this step are predominantly capillaries and small arterioles and venules, roughly 100 micrometers in diameter or smaller, which represent the functional workhorses of the cerebral circulation.

Once the vascular pellet has been collected, the protocol calls for rigorous washing of the isolated vessels over a cell strainer. This deceptively simple step removes trapped contaminants, including parenchymal cells and debris that can cling to the outside of vessel fragments and muddy downstream molecular profiles. The critical innovation, however, comes in the third stage: the optimized extraction of nuclei from the purified vessels using enzymatic digestion, specifically with collagenase III. The authors report that the precise conditions of this digestion step were tuned to liberate intact nuclei from endothelial, mural, and perivascular cells without compromising their quality, a balance that had defeated earlier attempts to profile vascular cells from frozen tissue.

After extraction, the protocol employs fluorescence-activated cell sorting, or FACS, to ensure that only high-purity nuclei proceed to sequencing. This sorting step is what makes the method compatible with the droplet-based platforms that dominate the field, including the widely used 10x Genomics single cell 3′ gene expression assay and the 10x multiome platform, which captures both gene expression and chromatin accessibility from the same nucleus. In parallel with the vascular nuclei, the protocol recovers parenchymal nuclei from the density gradient, meaning that a single dissection can yield both a vascular and a non-vascular view of the same tissue sample. Both fractions are purified by FACS, ensuring compatibility with single-nucleus RNA sequencing, single-nucleus ATAC sequencing, and combined multiomic workflows.

The practical demands of the protocol are modest by the standards of modern genomics. The authors estimate that the full workflow, from homogenized tissue to sorted nuclei ready for sequencing, requires approximately four to five hours to complete. Researchers with training in single-cell techniques and flow cytometry should be able to carry it out, which lowers the barrier to entry considerably compared with bespoke methods that demand specialized instrumentation or rare expertise. The protocol also accommodates both fresh and frozen tissue, a crucial feature for human research, where most brain samples arrive from brain banks as frozen material. Detailed guidance is provided for interpreting quality-control checkpoints along the way, including the use of Agilent BioAnalyzer traces during downstream library preparation, so that laboratories can diagnose and correct problems before committing precious samples to sequencing.

The significance of the method is best understood through the discoveries it has already enabled. VINE-seq formed the technical backbone of a human brain vascular atlas published in Nature in 2022, which revealed diverse mediators of Alzheimer’s disease risk expressed in specific vascular cell populations. Its multiome successor, MultiVINE-seq, extended the approach to paired measurements of gene expression and chromatin accessibility, and underpinned a 2025 study in Neuron demonstrating that human brain vascular multi-omics can elucidate disease-risk associations that transcriptomics alone cannot resolve. By publishing the complete protocol, the authors are effectively handing the wider research community the keys to a molecular map of the cerebrovasculature that was previously accessible only to a handful of specialist laboratories.

The broader implications reach into nearly every corner of neuroscience and neurology. The blood-brain barrier, formed by tightly joined endothelial cells and supported by pericytes and astrocyte endfeet, controls what enters the brain from the bloodstream and is implicated in systemic infection, inflammation, and the failure of countless drug candidates to reach their targets. Perivascular macrophages, which the protocol captures alongside endothelial and mural cells, are increasingly recognized as immunological sentinels with roles in neurodegeneration. Recent work has linked glycocalyx dysregulation to blood-brain barrier failure in aging, identified angiopoietin signaling as a central axis of amyloid-driven vascular dysfunction, and shown that depletion of the RNA-binding protein TDP-43 in endothelial cells disrupts core barrier pathways in neurodegeneration. Each of these lines of inquiry depends on the ability to molecularly profile vascular cells from real human tissue, precisely what VINE-seq and MultiVINE-seq make routine.

The authors have also made the underlying data openly available, with raw sequencing data for VINE-seq deposited in the NCBI Gene Expression Omnibus under accession GSE163577 and MultiVINE-seq data available in the Sequence Read Archive under BioProject PRJNA1182356. For a field that has spent more than half a century developing methods to isolate brain microvessels, dating back to pioneering capillary isolation work in the 1970s, the arrival of a standardized, hours-long workflow that feeds directly into single-nucleus and multiome sequencing marks a genuine inflection point. As laboratories around the world adopt the protocol, the vascular blind spot in brain atlases is likely to shrink rapidly, and with it, the gaps in our understanding of how the brain’s plumbing shapes neurological health, aging, and disease.

Subject of Research: Single-nucleus and multiome sequencing protocols for profiling cells of the brain vasculature

Article Title: VINE-seq and MultiVINE-seq for single-nucleus and multiome profiling of the brain vasculature

Article References: Oberhauser, J., Ding, B., Reid, M. M., Xie, W. H., & Yang, A. C. (2026). VINE-seq and MultiVINE-seq for single-nucleus and multiome profiling of the brain vasculature. Nature Protocols. https://doi.org/10.1038/s41596-026-01434-x

Image Credits: AI Generated

DOI: 10.1038/s41596-026-01434-x

Keywords: VINE-seq, MultiVINE-seq, brain vasculature, single-nucleus sequencing, blood-brain barrier, endothelial cells, pericytes, perivascular macrophages, FACS, Alzheimer's disease, multiome profiling, Nature Protocols