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Cited by 171RNA Interference and Gene DeliveryNanoparticle-Based Drug DeliveryBarrier Structure and Function Studies

Blood–brain-barrier-crossing lipid nanoparticles for mRNA delivery to the central nervous system

Wang, Chang, Xue, Yonger, Markovic, Tamara, Li, Haoyuan, Wang, Siyu, Zhong, Yichen, Du, Shi, Zhang, Yuebao +21 more·2025·Nature Materials
DOI 10.1038/s41563-024-02114-5PMC PMC12514551
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Abstract

The systemic delivery of mRNA molecules to the central nervous system is challenging as they need to cross the blood-brain barrier (BBB) to reach into the brain. Here we design and synthesize 72 BBB-crossing lipids fabricated by conjugating BBB-crossing modules and amino lipids, and use them to assemble BBB-crossing lipid nanoparticles for mRNA delivery. Screening and structure optimization studies resulted in a lead formulation that has substantially higher mRNA delivery efficiency into the brain than those exhibited by FDA-approved lipid nanoparticles. Studies in distinct mouse models show that these BBB-crossing lipid nanoparticles can transfect neurons and astrocytes of the whole brain after intravenous injections, being well tolerated across several dosage regimens. Moreover, these nanoparticles can deliver mRNA to human brain ex vivo samples. Overall, these BBB-crossing lipid nanoparticles deliver mRNA to neurons and astrocytes in broad brain regions, thereby being a promising platform to treat a range of central nervous system diseases.

AI snapshot

Written from the paper’s full text.

TL;DR

The authors synthesized 72 blood–brain-barrier-crossing lipids and identified MK16 lipid nanoparticles as a lead mRNA delivery platform for the central nervous system. MK16 BLNPs delivered functional mRNA to neurons and astrocytes across broad brain regions in mice and to human brain tissue ex vivo, with efficacy in cocaine-exposure and glioblastoma models.

Key findings

  • Screening 72 BBB-crossing lipids across six classes yielded MK16 BLNPs, which produced brain luminescence 8.3-fold greater than MC3 LNPs and 7.4-fold and 6.5-fold higher than ALC-0315 and SM-102 LNPs, respectively.
  • After intravenous GFP mRNA delivery at 0.5 mg kg−1, MK16 BLNPs transfected neurons (4.64 ± 1.00% GFP+), astrocytes (6.63 ± 0.62%), BCECs (5.62 ± 0.78%) and microglia (1.68 ± 0.15%); at 1 mg kg−1, these rose to 7.36 ± 0.78%, 9.71 ± 0.70%, 9.18 ± 0.80% and 2.85 ± 0.44%.
  • Cre mRNA delivery in Ai14 mice produced tdTomato in hippocampus, thalamus and cortex neurons (5.98 ± 1.52%, 5.86 ± 0.42%, 6.68 ± 1.13%) and astrocytes (8.36 ± 1.85%, 8.49 ± 1.53%, 8.74 ± 2.07%); triple injections increased tdTomato about twofold to 16.7 ± 2.2% in neurons, 19.2 ± 3.0% in astrocytes, 17.9 ± 1.6% in BCECs and 4.5 ± 0.9% in microglia.
  • BBB crossing involved caveolae- and γ-secretase-mediated transcytosis; MβCD reduced transcytosis by 43.3 ± 1.7% and MK-0752 by 63.8 ± 3.1%. In orthotopic U-118MG GBM, 70% of MK16 BLNP-Pten mRNA-treated mice survived over 120 days; in human cortical slices ex vivo, ΔFOSB was expressed in 4.00 ± 0.99% of neurons, 6.49 ± 2.33% of astrocytes and 2.38 ± 0.94% of microglia.

Methods

  • The authors conjugated BBB-crossing modules to amino lipids to create six classes of BBB-crossing lipids, formulated them with DOPE, cholesterol and DMG-PEG2k, and screened them in N2a and bEnd.3 cells before in vivo testing in C57BL/6J, Ai14 and NU/J mice.
  • Main assays included luciferase imaging, flow cytometry of brain cells, immunofluorescence, a bEnd.3 transwell BBB model with endocytosis inhibitors, RNA sequencing, blood biochemistry and histopathology, cocaine-induced conditioned place preference, orthotopic U-118MG GBM treatment, and ex vivo human cortical slice delivery.
  • Statistics used one-way ANOVA with Dunnett’s test, two-tailed Student’s t-test, two-way ANOVA with Šidák or Fisher’s LSD post hoc tests, and log-rank Mantel–Cox tests; P < 0.05 was considered significant.

Limitations

  • The paper states that future investigational new drug applications require good laboratory practice toxicology studies per FDA guidelines, including dose escalation and repeat administrations to uncover maximum-tolerated dose and potential long-term adverse effects, and that specific mRNA cargoes need careful design and examination.
  • Evident from the text: human validation was limited to ex vivo cortical tissue slices from two adult patients with Parkinson’s disease, with no in vivo human delivery or safety data.
  • Evident from the text: no statistical method was used to predetermine sample size, and many in vivo experiments used small groups (for example, n = 3 per group).

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1038/s41563-024-02114-5

Cites

57 of 57 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

20 of 28 cited sources checked · 3 supported · 3 flagged · accuracy not yet measured

  • Premisecitation

    d-Serine crosses the blood-brain barrier by amino acid transporter solute carrier family 6 member 14.

    “d-Serine serves as a neuromodulator and crosses the BBB by amino acid transporter solute carrier family 6 member 14 (refs. 31–35).”
    Quote in the paperSubstantial claim4 of 5 sources checked
    • cites [31] Hutchinson, K. et al. Describing inhibitor specificity for the amino acid transporter LAT1 from metainference simulations. Biophys. J. 121, 4476–4491 (2022). source

      The Introduction (page not provided) identifies LAT1 as SLC7A5 and says it is abundant at the blood-brain barrier. Searches for d-serine and SLC6A14 found no passage addressing whether d-serine crosses the barrier through SLC6A14. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [32] Wolosker, H., Blackshaw, S. & Snyder Solomon, H. Serine racemase: a glial enzyme synthesizing d-serine to regulate glutamate-N-methyl-d-aspartate neurotransmission. Proc. Natl Acad. Sci. USA 96, 13409–13414 (1999). source

      The Discussion describes D-serine as a modulator of NMDA neurotransmission, but does not address its passage across the blood-brain barrier. Searches for blood-brain barrier transport and SLC6A14 found no passage in our copy supporting the claimed transport mechanism. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [33] Bodner, O. et al. d-Serine signaling and NMDAR-mediated synaptic plasticity are regulated by system A-type of glutamine/d-serine dual transporters. J. Neurosci. 40, 6489 (2020). source

      The Introduction describes D-serine’s role in synaptic signaling, but does not discuss its passage across the blood–brain barrier. Searches for SLC6A14 and blood–brain barrier transport found no passage establishing the claimed route; the References instead list D-serine transport in the colon and a blood–brain barrier transporter for glutamine. (LLM, conf=0.77)

      Experimental — accuracy not yet measured
    • cites [34] Hatanaka, T. et al. Transport of d-serine via the amino acid transporter ATB0,+ expressed in the colon. Biochem. Biophys. Res. Commun. 291, 291–295 (2002). source

      The Results and Discussion identifies ATB0,+ as a transporter involved in intestinal absorption of d-serine, not transport across the blood–brain barrier. It instead attributes d-serine's access to the brain to system L1 (LAT1/4F2hc) at the blood–brain barrier. (LLM, conf=0.91)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured
    • cites [35] Ganapathy, M. E. & Ganapathy, V. Amino acid transporter ATB0,+ as a delivery system for drugs and prodrugs. Curr. Drug Targets: Immune, Endocr. Metab. Disord. 5, 357–364 (2005). source
  • Premisecitation

    Increasing the ionizable lipid molar ratio in an LNP formulation can augment mRNA delivery.

    “In our prior studies, we found that the increase in ionizable lipid molar ratio in the LNP formulation can augment mRNA delivery44.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [44] Li, B. et al. An orthogonal array optimization of lipid-like nanoparticles for mRNA delivery in vivo. Nano Lett. 15, 8099–8107 (2015). source

      In the formulation-optimization passage (labeled Abstract; page unavailable), the paper says that reduced levels of the lipid-like component TT3 may facilitate mRNA delivery and describes decreasing its ratio from 40 to 10. It does not report increased delivery from increasing the TT3 ratio; its reported trend runs in the opposite direction. (LLM, conf=0.84)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured
  • Premisecitation

    Delivery of mRNA to the brain remains a formidable challenge due to the blood-brain barrier.

    “However, the delivery of mRNA to the brain remains a formidable challenge due to the blood–brain barrier (BBB)18,19.”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [18] Xiong, B. et al. Strategies for structural modification of small molecules to improve blood–brain barrier penetration: a recent perspective. J. Med. Chem. 64, 13152–13173 (2021). source
    • cites [19] Brown, K. M. et al. Expanding RNAi therapeutics to extrahepatic tissues with lipophilic conjugates. Nat. Biotechnol. 40, 1500–1508 (2022). source

      The Introduction and Discussion (pages unavailable) describe the blood–brain barrier as a challenge for delivering siRNA to the CNS, supporting the general barrier point. The paper addresses siRNA, not mRNA, so it does not establish the claim specifically for mRNA delivery. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
  • Premisecitation

    Tryptamine derivatives cross the blood-brain barrier by active transport via Mg2+ and ATP-dependent uptake.

    “Tryptamine derivatives are neurotransmitter precursors that cross the BBB by active transport via Mg2+ and ATP-dependent uptake38–40.”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [38] Carbonaro, T. M. & Gatch, M. B. Neuropharmacology of N,N-dimethyltryptamine. Brain Res. Bull. 126, 74–88 (2016). source

      Section 2.2, “Accumulation and storage,” says that DMT crosses the blood–brain barrier by active transport involving Mg2+- and ATP-dependent uptake. It does not establish that this mechanism applies to tryptamine derivatives generally. Section 2.1 describes tryptamine as a precursor to DMT, but does not characterize tryptamine derivatives as neurotransmitter precursors. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [39] Ma, F. et al. Neurotransmitter-derived lipidoids (NT-lipidoids) for enhanced brain delivery through intravenous injection. Sci. Adv. 6, eabb4429 (2020). source

      The Introduction (page unavailable) says that dimethyltryptamine and other tryptamine derivatives have been shown to cross the BBB by active transport. It does not specify Mg2+- or ATP-dependent uptake; searches for those mechanisms found no supporting passage in our copy. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [40] Nagai, F., Nonaka, R. & Satoh Hisashi Kamimura, K. The effects of non-medically used psychoactive drugs on monoamine neurotransmission in rat brain. Eur. J. Pharmacol. 559, 132–137 (2007). source
  • Premisecitation

    l-DOPA is recognized by the large neutral amino acids transporter 1 on endothelial cells and transported across the blood-brain barrier for treatment of Parkinson's disease.

    “Specifically, l-DOPA can be recognized by the large neutral amino acids transporter 1 on endothelial cells and be transported across the BBB for the treatment of Parkinson’s disease29–31.”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [29] Abbott, A. Levodopa: the story so far. Nature 466, S6–S7 (2010). source

      The opening section (p. 1) describes l-DOPA as a dopamine precursor that crosses the blood-brain barrier to treat Parkinson’s disease. The paper does not identify large neutral amino acid transporter 1 or endothelial cells as the mechanism of transport. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [30] Kageyama, T. et al. The 4F2hc/LAT1 complex transports l-DOPA across the blood–brain barrier. Brain Res. 879, 115–121 (2000). source
    • cites [31] Hutchinson, K. et al. Describing inhibitor specificity for the amino acid transporter LAT1 from metainference simulations. Biophys. J. 121, 4476–4491 (2022). source

      The Introduction (page not provided) states that LAT1 is abundant in blood-brain-barrier endothelial cells and mediates levodopa (L-DOPA) transport across the barrier. It does not state that this transport is for treatment of Parkinson’s disease; that treatment detail is not established by the eligible Introduction passages. (LLM, conf=0.78)

      Experimental — accuracy not yet measured
  • Classified as probably not a substantial claim. 14
  • May combine two claims. 27

  • Probably not a substantial claim: may not pass S1 (asserted). May combine two claims.

    Premisecitation

    Enhancing ΔFOSB could be a viable approach to treat drug addiction by making subthreshold, non-reinforcing doses of cocaine more reinforcing, potentially reducing the risks of overdose.

    “Given that the major clinical problem in addiction is that the user becomes tolerant to the effects of the drug over time, prior studies reported that enhancing ΔFOSB could be a viable approach to treat drug addiction by making subthreshold, non-reinforcing doses of cocaine more reinforcing, potentially reducing the risks of overdose51.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [51] Zhang, Y. et al. Overexpression of deltaFosB in nucleus accumbens mimics the protective addiction phenotype, but not the protective depression phenotype of environmental enrichment. Front. Behav. Neurosci. 8, 297 (2014). source

      The Results (page unavailable in our copy) say that differences in cocaine intake appeared only at higher doses, not at the low doses the claim says became more reinforcing. The Discussion reports that ΔFosB overexpression in these rats decreased cocaine intake and seeking; it contrasts this with earlier findings of increased sensitivity to low-dose cocaine reward in transgenic mice. The paper does not propose reduced overdose risk through the mechanism claimed. (LLM, conf=0.86)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured

  • Classified as probably not a substantial claim. May combine two claims.

    Premisecitation

    Temozolomide is an alkylating lipophilic agent that crosses the blood-brain barrier to treat GBM.

    “Temozolomide is an alkylating lipophilic agent that crosses the BBB to treat GBM36,37.”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites [36] Darlix, A. et al. Chemotherapy and diffuse low-grade gliomas: a survey within the European Low-Grade Glioma Network. Neurooncol. Pract. 6, 264–273 (2019). source

      The Abstract (Conclusions; page unavailable) identifies temozolomide as a chemotherapy drug, and the Discussion (Choice of Chemotherapy Regimen; page unavailable) reports its use and efficacy in diffuse low-grade gliomas. The paper does not establish that it is lipophilic or alkylating, crosses the blood–brain barrier, or is used to treat GBM; those parts of the claim are unsupported by this copy. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
    • cites [37] de Gooijer, M. C. et al. Improved brain penetration and antitumor efficacy of temozolomide by inhibition of ABCB1 and ABCG2. Neoplasia 20, 710–720 (2018). source

      The Introduction identifies temozolomide as an alkylating agent used in standard GBM treatment and says it penetrates the blood-brain barrier. The Discussion also reports that it readily permeates membranes, but our copy does not establish the specific description “lipophilic.” (LLM, conf=0.86)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Cinnamic derivatives can cross the blood-brain barrier and insert into the beta sheet of amyloid β proteins.

    “Cinnamic derivatives can cross the BBB and insert into the beta sheet of amyloid β proteins41.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [41] Yang, J. et al. Turn-on chemiluminescence probes and dual-amplification of signal for detection of amyloid beta species in vivo. Nat. Commun. 11, 4052 (2020). source

      The Results (In vivo imaging with ADLumin-1; page unavailable) report that ADLumin-1 crossed the blood–brain barrier in mice. The Results (Design and synthesis of ADLumin-1; page unavailable) say its binding moiety has the potential to insert into Aβ beta sheets, but the paper does not identify the probes as cinnamic derivatives or establish insertion as a demonstrated fact. (LLM, conf=0.84)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductcitation

    An unbiased CPP test was carried out as described in reference 52.

    “An unbiased CPP test was carried out as described52”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [52] Robison, A. J. et al. Behavioral and structural responses to chronic cocaine require a feedforward loop involving ΔFosB and calcium/calmodulin-dependent protein kinase II in the nucleus accumbens shell. J. Neurosci. 33, 4295–4307 (2013). source

      The Materials and Methods section describes locomotor-response tests (Experiments 2 and 16), but the retrieved passages do not describe an unbiased conditioned place preference (CPP) test. Searches for “CPP,” “place preference,” and an unbiased conditioning protocol did not locate the claimed method in our copy. (LLM, conf=0.74)

      Experimental — accuracy not yet measured

  • Classified as probably not a substantial claim.

    Premisecitation

    ΔFOSB is a pivotal transcription factor involved in the regulation of addictive behaviours and the neuropathology of drug dependence.

    “ΔFOSB is a pivotal transcription factor involved in the regulation of addictive behaviours and the neuropathology of drug dependence49,50.”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [49] Kelz, M. B. et al. Expression of the transcription factor ΔFosB in the brain controls sensitivity to cocaine. Nature 401, 272–276 (1999). source
    • cites [50] Nestler, E. J., Barrot, M. & Self, D. W. ΔFosB: a sustained molecular switch for addiction. Proc. Natl Acad. Sci. USA 98, 11042–11046 (2001). source

  • May combine two claims.

    Premisecitation

    PTEN is commonly mutated in wide ranges of GBM, which may be a candidate for therapeutic intervention.

    “A tumour suppressor gene, like phosphatase and tensin homologue (PTEN), is commonly mutated in wide ranges of GBM, which may be a great candidate for therapeutic intervention27,54.”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites [27] Yang, Z. et al. Large-scale generation of functional mRNA-encapsulating exosomes via cellular nanoporation. Nat. Biomed. Eng. 4, 69–83 (2020). source

      The Results section ('Functional and pharmacokinetic evaluation of CNP generated exosomes'; page not specified) calls PTEN a commonly mutated tumour suppressor gene and tests it as a therapeutic target in two PTEN-deficient glioma models. The in vivo Results report inhibited tumour growth after PTEN-restoring treatment. The copy does not establish how widely PTEN is mutated specifically across GBM. (LLM, conf=0.83)

      Experimental — accuracy not yet measured
    • cites [54] Hopkins, B. D., Hodakoski, C., Barrows, D., Mense, S. M. & Parsons, R. E. PTEN function: the long and the short of it. Trends Biochem. Sci. 39, 183–190 (2014). source

      The ‘Importance of PTEN function and regulation’ section (page not provided) identifies PTEN as a tumour suppressor frequently lost in cancers including brain tumours and reports mutations across many tumour types. It does not establish how commonly PTEN is mutated specifically in GBM or identify it as a therapeutic candidate for GBM. (LLM, conf=0.85)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Findingown data

    BLNPs have particle sizes ranging from 97.3 ± 2.0 nm to 177.6 ± 2.0 nm with a polydispersity index of <0.3.

    “The particle sizes of BLNPs ranged from 97.3 ± 2.0 nm to 177.6 ± 2.0 nm with a polydispersity index of <0.3 (Supplementary Fig. 1a,b).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    After intravenous injection of MK16-Alexa 647 RNA BLNPs, nearly 56.6 ± 2.5% of the MK16 BLNPs were in the liver, 6.5 ± 1.1% in the spleen, 7.4 ± 0.8% in the kidneys, 10.5 ± 1.0% in the lungs and 3.7 ± 1.4% in the heart.

    “Using this method, we observed nearly 56.6 ± 2.5% of the MK16 BLNPs in the liver, 6.5 ± 1.1% in the spleen, 7.4 ± 0.8% in the kidneys, 10.5 ± 1.0% in the lungs and 3.7 ± 1.4% in the heart (Supplementary Fig. 10b).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    MK16 BLNPs exhibited a particle size of 137.0 ± 4.1 nm and an mRNA encapsulation efficiency of approximately 84.8 ± 1.5%.

    “The MK16 BLNPs exhibited a particle size of 137.0 ± 4.1 nm and an mRNA encapsulation efficiency of approximately 84.8 ± 1.5% (Supplementary Fig. 9a,b).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    MK6 BLNPs induced the highest luminescence intensity in brain tissue 6 h after intravenous administration among all BLNPs tested, much higher than MC3 LNPs.

    “Among all the BLNPs tested, MK6 BLNPs induced the highest luminescence intensity in brain tissue 6 h after administration, which was much higher than MC3 LNPs (Fig. 2g,h).”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Findingown data

    MK16 BLNPs were positively charged and displayed spherical morphology in cryogenic transmission electron microscopy images.

    “The particles were positively charged and displayed spherical morphology in cryogenic transmission electron microscopy images (Supplementary Fig. 9c).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    MK16 BLNPs showed potent mRNA delivery efficiency in both bEnd.3 cells and N2a cells with negligible cytotoxicity at the tested dose.

    “Additionally, MK16 BLNPs showed potent mRNA delivery efficiency in both bEnd.3 cells and N2a cells with negligible cytotoxicity at the tested dose (Supplementary Fig. 9d,e).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    MK16 BLNPs preferentially delivered GFP mRNA to neurons (4.64 ± 1.00% GFP+), astrocytes (6.63 ± 0.62% GFP+) and brain capillary endothelial cells (5.62 ± 0.78% GFP+) at an mRNA dose of 0.5 mg kg−1, superior to MC3 LNPs in corresponding cell types.

    “MK16 BLNPs preferentially delivered GFP mRNA to neurons (4.64 ± 1.00% GFP+), astrocytes (6.63 ± 0.62% GFP+) and brain capillary endothelial cells (BCECs; 5.62 ± 0.78% GFP+) at an mRNA dose of 0.5 mg kg−1, the efficiency of which is particularly superior to those achieved by MC3 LNPs in the corresponding cell types (Fig. 3f,g and Supplementary Fig. 11b).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Approximately 83.1 ± 2.7% of mRNA was encapsulated in MK6 BLNPs, and they were slightly positively charged.

    “Approximately 83.1 ± 2.7% of mRNA was encapsulated in MK6 BLNPs, and they were slightly positively charged (Supplementary Fig. 5c).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    MK16 BLNPs showed the highest in vivo luminescence intensity in the mouse brain after intravenous injection among the MK series, 1.7-fold higher than MK6E BLNPs and 8.3-fold greater than MC3 LNPs.

    “Among this series of MK BLNPs, MK16 BLNPs showed the highest in vivo luminescence intensity in the mouse brain after intravenous injection, which was 1.7-fold higher than MK6E BLNPs and 8.3-fold greater than MC3 LNPs (Fig. 3d,e).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    MK6 BLNPs were around 139.8 ± 3.5 nm in diameter with a polydispersity index of <0.15.

    “MK6 BLNPs were around 139.8 ± 3.5 nm in diameter with a polydispersity index of <0.15 (Supplementary Fig. 5b).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    MK16 BLNPs showed 7.4-fold and 6.5-fold higher luminescence intensity than ALC-0315 and SM-102 LNPs in brains after intravenous administration, respectively.

    “Additionally, MK16 BLNPs showed 7.4-fold and 6.5-fold higher luminescence intensity than ALC-0315 and SM-102 LNPs in brains after intravenous administration, respectively (Supplementary Fig. 8a,b).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Substantial levels of fluorescent signals from MK16 BLNPs loaded with Alexa Fluor 647-labelled RNA were observed in the brain at 1 h and 6 h after a single intravenous injection, with no detectable signals at and after 18 h.

    “Substantial levels of fluorescent signals were observed in the brain at the time points of 1 h and 6 h, with no detectable signals at and after the 18 h time point (Supplementary Fig. 10a).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    MK16 BLNP-ΔFosb mRNA treatment resulted in ΔFOSB expression in 4.00 ± 0.99% of neurons, 6.49 ± 2.33% of astrocytes and 2.38 ± 0.94% of microglia in adult human cortical tissue ex vivo, whereas untreated slices exhibited no detectable expression.

    “Quantitative analysis revealed ΔFOSB expression in 4.00 ± 0.99% of neurons, 6.49 ± 2.33% of astrocytes and 2.38 ± 0.94% of microglia, whereas the untreated slices exhibited no detectable expression (Fig. 5e).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    MK16 BLNP-Pten mRNA reduced tumour growth and prolonged survival in tumour-bearing mice compared with PBS and MK16 BLNP-control mRNA (mCherry mRNA) groups.

    “Moreover, MK16 BLNP-Pten mRNA reduced the tumour growth, thereby achieving prolonged survival in tumour-bearing mice compared with the PBS and MK16 BLNP-control mRNA (mCherry mRNA) groups (Fig. 6b–d).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    On MK16 BLNP delivery of Cre mRNA, a tdTomato signal was observed throughout most regions of the brain, with the strongest signal detected in brain margins.

    “In PBS-treated mice, tdTomato expression was undetectable 5 days after administration, but on MK16 BLNP delivery of Cre mRNA, a tdTomato signal was observed throughout most regions of the brain, with the strongest signal detected in brain margins (Fig. 4b and Supplementary Fig. 13a,b).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In a mouse GBM model, MK16 BLNP-Pten mRNA treatment significantly reduced tumour growth and enhanced survival rates compared with MC3 LNP-Pten mRNA treatment.

    “mice treated with MK16 BLNP-Pten mRNA showed significantly reduced tumour growth and enhanced survival rates compared with those receiving MC3 LNP-Pten mRNA (Supplementary Fig. 19a–c).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In the hippocampus, tdTomato-positive neurons in the MK16 BLNP group accounted for 5.98 ± 1.52% of the total; in the thalamus and cortex, the proportions were 5.86 ± 0.42% and 6.68 ± 1.13%, respectively.

    “Specifically, in the hippocampus, tdTomato-positive neurons in the MK16 BLNP group accounted for 5.98 ± 1.52% of the total, whereas in the thalamus and cortex, the proportions were 5.86 ± 0.42% and 6.68 ± 1.13%, respectively (Fig. 4f–h).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Astrocytes with the tdTomato signal represented 8.36 ± 1.85% in the hippocampus, 8.49 ± 1.53% in the thalamus and 8.74 ± 2.07% in the cortex.

    “Astrocytes with the tdTomato signal represented 8.36 ± 1.85% in the hippocampus, 8.49 ± 1.53% in the thalamus and 8.74 ± 2.07% in the cortex (Fig. 4f–h).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    Both BBB-crossing modules and diverse structures of amino lipids can greatly affect the in vitro mRNA delivery efficiency.

    “These results suggest that both BBB-crossing modules and diverse structures of amino lipids can greatly affect the in vitro mRNA delivery efficiency.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Several approaches can mediate crossing of the blood-brain barrier, including passive diffusion and adsorptive-mediated, carrier-mediated and receptor-mediated transcytosis.

    “Previous studies have reported that several approaches can mediate the crossing of the BBB, including passive diffusion and adsorptive-mediated, carrier-mediated and receptor-mediated transcytosis25.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [25] Terstappen, G. C., Meyer, A. H., Bell, R. D. & Zhang, W. Strategies for delivering therapeutics across the blood–brain barrier. Nat. Rev. Drug Discov. 20, 362–383 (2021). source

  • May combine two claims.

    Interpretationown data

    MK16 BLNPs can cross endothelial cells by both caveolae- and γ-secretase-mediated transcytosis.

    “These results indicate that MK16 BLNPs can cross endothelial cells by both caveolae- and γ-secretase-mediated transcytosis.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductnone given

    To mitigate any pre-existing bias towards a particular chamber, the groups were carefully balanced.

    “To mitigate any pre-existing bias towards a particular chamber, the groups were carefully balanced.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductreasoning

    All relevant ethical regulations were followed as applicable.

    “All relevant ethical regulations were followed as applicable.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S2 (checkable). May combine two claims.

    Interpretationreasoning

    The BLNP platform represents an important proof-of-concept study to advance LNP-mRNA delivery to the brain, offering a possible avenue for treating a wide range of brain disorders.

    “In summary, this BLNP platform represents an important proof-of-concept study to advance LNP-mRNA delivery to the brain, which offers a possible avenue for treating a wide range of brain disorders.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    The mice were divided into three random groups: PBS group, control (mCherry) mRNA group and Pten mRNA group.

    “The mice were divided into three random groups: PBS group, control (mCherry) mRNA group and Pten mRNA group.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S4 (based). May combine two claims.

    Study design and conductnone given

    All the data met the assumptions of the statistical tests used, including normality and equal variances, and were formally tested to ensure the validity of the statistical analysis.

    “All the data met the assumptions of the statistical tests used, including normality and equal variances, and were formally tested to ensure the validity of the statistical analysis.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    MK16 BLNPs have biocompatibility and safety.

    “These results highlight the biocompatibility and safety of MK16 BLNPs.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductnone given

    All research involving human subjects was conducted under STUDY-13-00415 of the Human Research Protection Program at the Icahn School of Medicine at Mount Sinai.

    “All research involving human subjects was conducted under STUDY-13–00415 of the Human Research Protection Program at the Icahn School of Medicine at Mount Sinai.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductnone given

    No statistical method was used to predetermine the sample size.

    “No statistical method was used to predetermine the sample size.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductnone given

    The animal experiments conducted in this study adhered to the guidelines approved by the Institutional Animal Care and Use Committee of the Ohio State University and the Icahn School of Medicine at Mount Sinai.

    “The animal experiments conducted in this study adhered to the guidelines approved by the Institutional Animal Care and Use Committee of the Ohio State University (2014A00000106) and the Icahn School of Medicine at Mount Sinai (IPROTO202200000134).”
    Quote in the paperNot a substantial claim
36 of 36

  • The paper relies on this without support.

    There is an urgent demand to effectively and safely deliver mRNA to brain cells, especially neurons.

    “Despite these significant advances, there is an urgent demand to effectively and safely deliver mRNA to brain cells, especially neurons.”

  • The authors state this as an assumption.

    Conjugating BBB-crossing modules to amino lipids yields BBB-crossing lipids that can facilitate blood–brain barrier crossing of the resulting lipid nanoparticles.

    “On the basis of these important findings, we designed and synthesized six classes of BBB-crossing lipids (BLs) by conjugating these molecules with various amino lipids. We hypothesize that these BLs can facilitate formulations of BBB-crossing lipid nanoparticle (BLNP) with mRNA that cross the BBB and efficiently deliver mRNA to the central nervous system (Fig. 1a).”
2 of 2

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Six series of brain-targeting lipids (BLs) were previously designed for efficient mRNA delivery to the brain through intravenous administration.

    “We previously designed and developed six series of ionizable lipids referred to as brain-targeting lipids (BLs), for efficient mRNA delivery to the brain through intravenous administration.[13]”

    From Lipid Nanoparticles Enhance mRNA Delivery to the Central Nervous System Upon Intrathecal Injection (2025) · cites this paper as [13]

Cites

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3 of the 60 references couldn’t be matched to a known paper.

Cited by

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Related

Authors
Wang, C., Xue, Y., Markovic, T., Li, H., Wang, S., Zhong, Y., Du, S., Zhang, Y., Hou, X., Yu, Y., Liu, Z., Tian, M., Kang, D. D., Wang, L., Guo, K., Cao, D., Yan, J., Deng, B., McComb, D. W., Parsons, R. E., Minier-Toribio, A. M., Holt, L. M., Pan, J., Hashemi, A., Kopell, B. H., Charney, A. W., Nestler, E. J., Peng, P. C., & Dong, Y.
Affiliations
  • Tisch Cancer Institute
  • Icahn School of Medicine at Mount Sinai (US)
  • The Ohio State University (US)
  • Biogen (United States) (US)
Corresponding author
  • Nestler, Eric J.
  • Peng, Paul C.
  • Dong, Yizhou
ORCID
Venue
Nature Materials
Publisher
Springer Science and Business Media LLC
Published
2025-02-17
Volume
24
Issue
10
Pages
1653-1663
ISSN
  • 1476-1122 (ISSN-L)
  • 1476-4660
DOI
10.1038/s41563-024-02114-5
PMC
PMC12514551
OpenAlex
W4407661809
Cited by
171
References
60
Citations by year
2026: 106 · 2025: 65
FWCI
42.91
Type
journal article
Language
en
Open access
closed
Licence
Locations
  • Nature Materials (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (submittedVersion)
Funders
  • U.S. Department of Health & Human Services | NIH | National Institute of General Medical Sciences (R35GM144117)
  • U.S. Department of Health & Human Services | NIH | National Institute on Drug Abuse (P01DA047233)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
  • Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
  • Life Sciences › Neuroscience › Neurology › Barrier Structure and Function Studies
Keywords
blood-brain barrier penetration, lipid nanoparticles, mRNA delivery, central nervous system, intravenous delivery, neuronal transfection, central nervous system disorders
MeSH terms
Animals, Astrocytes, Blood-Brain Barrier, Brain, Central Nervous System, Humans, Lipids, Liposomes, Neurons, RNA, Messenger, Mice, Nanoparticles

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-20.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-20.

Cited sources are checked by gpt-6-sol.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

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