Illustration of CNF-sheet-mediated membrane fusion between an extracellular vesicle and a lipid nanoparticle carrying molecular beacons for internal microRNA detection.
Image Credit: Courtesy of Nagoya University and the researchers
Scientific Frontline: Extended "At a Glance" Summary: Single-Vesicle Colocalization Sensing
The Core Concept: Researchers have developed a novel, non-destructive diagnostic technique that analyzes both the surface proteins and internal microRNAs of individual extracellular vesicles in blood and urine. By linking internal molecular cargo to specific surface markers without breaking the vesicle membrane, this method identifies the precise tissue originating chronic inflammation.
Key Distinction/Mechanism: Conventional methods require breaking open extracellular vesicle membranes to measure internal microRNAs, which mixes molecules from multiple vesicles and severs the link to their original surface proteins. This new approach utilizes cellulose nanofiber sheets to capture the vesicles and promotes membrane fusion with lipid nanoparticles carrying molecular beacons; these beacons enter the intact vesicle and fluoresce upon binding to specific target microRNAs, allowing simultaneous reading of internal cargo and tissue-origin surface markers.
Origin/History: This technique was developed by a research team at Nagoya University, with the findings published in Cell Reports Physical Science in 2026.
Major Frameworks/Components:
- Extracellular Vesicles: Membrane-bound particles released by cells that carry molecular information, including microRNAs and proteins, throughout body fluids.
- Cellulose Nanofiber Sheets: Wood-derived, nanoscale mesh structures utilized to capture, retain, and fuse membrane-bound particles during the drying process.
- Lipid Nanoparticle-Delivered Molecular Beacons: Probes delivered inside vesicles via membrane fusion that emit fluorescence when bound to target microRNAs, enabling non-destructive internal analysis.
- Colocalization Biomarkers: The specific pairing of general inflammatory microRNAs, such as miR-1246, with tissue-specific surface proteins, such as DPYS for the kidney, to trace the anatomical source of an immune response.
Branches of Science: Biomolecular Engineering, Cell Biology, Molecular Biology, Molecular Pathology.
Future Application: This methodology enables the development of non-invasive liquid biopsy tests using blood or urine to evaluate and monitor chronic inflammatory diseases, potentially accelerating clinical interventions for organ rejection, autoimmune diseases, and brain disorders.
Why It Matters: Chronic inflammation underlies numerous severe diseases and causes irreversible tissue damage over time, yet traditional biomarkers, such as C-reactive protein, cannot identify the specific affected organ. This innovation allows clinicians to pinpoint the exact anatomical source of prolonged immune responses without relying on invasive tissue biopsies.
Developed at Nagoya University, a new technique to diagnose chronic inflammation can read information from the surface and within individual extracellular vesicles in blood and urine. This technique could be used to develop noninvasive tests that not only detect chronic inflammation but also provide information about the tissues where it occurs.
Whenever the body faces damage from injury, illness, or toxins, it launches an inflammatory response that dilates blood vessels to rush fluid and nutrients to the affected site and sends inflammatory cells to destroy pathogens and repair tissue. However, the immune system can sometimes continue sending inflammatory cells to attack tissues or organs over months or years, even in the absence of any threat. Known as chronic inflammation, it underlies cancers, brain disorders, autoimmune diseases, and heart disease, gradually causing irreversible tissue damage. Early detection is critical because prolonged inflammation contributes to disease progression.
“Currently, there are several conventional inflammatory biomarkers, such as C-reactive protein,” said Taiga Ajiri, an assistant professor at Nagoya University’s Yasui Lab, part of the Department of Biomolecular Engineering. “These biomarkers can detect inflammation, but it is difficult to identify where it occurs.”
To locate inflammation, researchers turned to extracellular vesicles (EVs)—tiny, membrane-bound particles released by cells throughout the body that carry molecular information from their cells of origin. EVs function like parcels with tracking labels. Proteins on their membranes provide clues about their tissue of origin, while microRNAs inside EVs reflect the condition of the originating cells. The vesicle membrane protects this cargo as it travels through body fluids, such as blood and urine.
Conventionally, measuring internal microRNAs requires breaking open EV membranes, mixing molecules from many vesicles together. This makes it difficult to link the internal information to surface proteins. Reading both internal microRNA and surface protein information without breaking the EV membrane, however, could reveal previously hidden EV subpopulations.
Reading an EV Package
The researchers used cellulose nanofiber (CNF) sheets made from wood-derived cellulose. The sheets' mesh-like nanoscale pores capture and retain EVs. Previous work by Professor Takao Yasui’s group established the use of these sheets for EV capture and analysis.
The team found that the drying process on CNF sheets brings membrane-bound particles into close contact, promoting membrane fusion. They applied this property to combine EVs with lipid nanoparticles containing molecular beacons—probes that detect specific microRNAs.
“These lipid nanoparticles can deliver molecular beacons inside the EV without disrupting the EV,” Ajiri said. These beacons emit fluorescence when they bind to specific microRNAs, allowing researchers to detect the presence of target molecules.
Clues to the Source: Kidney Inflammation
To test the method, the team established a laboratory model of chronic active antibody-mediated rejection (CAAMR), a complication of kidney transplantation, by exposing human kidney cells to inflammatory signals. They identified seventy-one microRNAs with altered expression in released EVs and focused on the highly abundant miR-1246. They also selected DPYS, a kidney-associated membrane protein, to combine information on inflammation with information on tissue origin.
After accounting for detection efficiency, the researchers estimated that about one in ten EVs from unstimulated kidney cells contained miR-1246, compared to about four in ten after inflammatory stimulation. By contrast, quantitative PCR (qPCR) analysis using equivalent sample inputs did not detect a statistically significant difference.
The team then analyzed urine from kidney transplant patients with CAAMR, patients with immune diseases not primarily involving the kidney, and healthy volunteers.
Neither DPYS nor miR-1246 alone clearly distinguished the patient groups. EVs carrying both miR-1246 and the general EV marker CD63 increased in inflammatory disease groups, but this combination could not distinguish renal from nonrenal inflammation. In contrast, EVs carrying both DPYS and miR-1246 were enriched in CAAMR patients, providing clues to inflammation associated with the transplanted kidney.
The team hopes to improve particle recovery and streamline the CNF-based procedure while testing the approach in larger clinical studies. Because urine and blood can be collected with relatively little burden, the method may contribute to less-invasive evaluation and monitoring of chronic inflammatory diseases.
Reference material: What Is: Extracellular Vesicles (Exosomes)
Funding: This research was supported by the Moonshot Research and Development Program (grant nos. 22zf0127004s0902 and JP22zf0127009) from the Japan Agency for Medical Research and Development (AMED) (to T. Yasui); AMED (JP243fa627005, JP21zf0127004, and JP23gm1910005 to M. Murakami; 24ae0121056 to H.A.); the New Energy and Industrial Technology Development Organization (NEDO) JPNP20004 (to T. Yasui and H.K.); Q-Leap (JPMXS0120330644 to M. Murakami); the Japan Science and Technology Agency (JST) CREST (JPMJCR2576 to T. Yasui), AIP Acceleration Research (JPMJCR23U1 to T. Yasui), and FOREST (JPMJFR2003 to H.K.); the JSPS Grant-in-Aid for Scientific Research (A) 24H00792 (to T. Yasui); the JSPS Grant-in-Aid for Scientific Research (S) 21H05025 (to H.A.); the JSPS Grant-in-Aid for Scientific Research (S) 24H00038 (to M.T.); the JSPS Grant-in-Aid for Scientific Research (B) 23K28408 (to M. Maeki); the JSPS Grant-in-Aid for Scientific Research (A) 23H00541 (to M. Maeki); the JSPS Grant-in-Aid for Transformative Research Areas (B) 25H01414 (to Y.K.); JSPS Grant-in-Aid for Young Scientists (Start-up) 24K23101 (to T.A.); Platform Project for Supporting Drug Discovery and Life Science Research (Basis for Supporting Innovative Drug Discovery and Life Science Research [BINDS]) from AMED under grant no. JP24ama121038 (support no. 4008) (to T. Yokoyama); Grant for Joint Research Program of the Institute for Genetic Medicine, Hokkaido University, by the Photo-excitonic Project, Hokkaido University, and by the Promotion Project for Young Investigators at Hokkaido University, Japan (to M. Murakami); Science Tokyo SPRING grant no. KH240019nn (to S.K.); JSPS Research Fellowship 25KJ1250 (to S.K.); and the Noguchi Institute NJ202308 (to T. Yasui).
Published in journal: Cell Reports Physical Science
Authors: Shota Kawaguchi, Taiga Ajiri, Kunanon Chattrairat, Hirotaka Koga, Yukiko Kamiya, Fuminori Sato, Hiroyuki Asanuma, Masatoshi Maeki, Manabu Tokeshi, Takeshi Yokoyama, Yoshikazu Tanaka, Kosuke Yoshida, Akira Yokoi, Kiyohiko Hotta, Naoya Iwahara, Yuichiro Fujieda, Mitsutaka Yasuda, Tatsuya Atsumi, Shimpei I. Kubota, Kaoru Murakami, Masaaki Murakami, and Takao Yasui
Source/Credit: Nagoya University
Edited by: Scientific Frontline
Reference Number: cbio110226_01
