Designing and optimizing AAV-mediated gene therapy for neurodegenerative diseases: from bench to bedside

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

The most commonly used ubiquitous promoters include CAG, CBA, and CMV promoters.

“Among the most commonly used ubiquitous promoters are cytomegalovirus immediate early enhancer/chicken β actin/rabbit β-globin (CAG) promoter, the chicken beta-acting (CBA) promoter and the cytomegalovirus (CMV) promoter [18].”

• cites [18] Au HKE, Isalan M, Mielcarek M. Gene therapy advances: a meta-analysis of AAV usage in clinical settings. Front Med (Lausanne). 2021;8: 809118. sourceSupported

May combine two claims.

Intron elements are generally positioned downstream of a promoter and enhance gene expression by improving mRNA stability and facilitating mRNA export to the cytoplasm.

“Intron element is generally positioned downstream of a promoter and enhances gene expression by improving mRNA stability and facilitating mRNA export to the cytoplasm [23].”

• cites [23] Reed R, Hurt E. A conserved mRNA export machinery coupled to pre-mRNA splicing. Cell. 2002;108(4):523–31. sourcePartly supported

  The passage describing the splicing-coupled export model (labelled Abstract; page not supplied) says that splicing recruits export factors and that the spliced mRNA moves to the cytoplasm. It does not say that intron elements are generally positioned downstream of a promoter or that they enhance gene expression by improving mRNA stability. (LLM, conf=0.84)

  Experimental — accuracy not yet measured

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

In silico design approach to reconstruct and discover new capsid was first reported in 2015.

“With technological advancements of computational modeling and machine learning, an in silico design approach to reconstruct and discover new capsid was first reported in 2015 [69, 70].”

• cites [69] Zinn E, Pacouret S, Khaychuk V, Turunen HT, Carvalho LS, Andres-Mateos E, Shah S, Shelke R, Maurer AC, Plovie E, et al. In silico reconstruction of the viral evolutionary lineage yields a potent gene therapy vector. Cell Rep. 2015;12(6):1056–68. sourcePartly supported

  The Discussion (page unavailable) says the authors inferred ancestral AAV capsid sequences through in silico modeling and identified a new AAV-like particle, Anc80L65. It does not establish that this approach was first reported in 2015; it describes an earlier viral-ancestor reconstruction as a methodological precedent. (LLM, conf=0.86)

  Experimental — accuracy not yet measured
• cites [70] Santiago-Ortiz J, Ojala DS, Westesson O, Weinstein JR, Wong SY, Steinsapir A, Kumar S, Holmes I, Schaffer DV. AAV ancestral reconstruction library enables selection of broadly infectious viral variants. Gene Ther. 2015;22(12):934–46. sourcePartly supported

  The Introduction describes reconstructing the AAV capsid and selecting new, broadly infectious variants, supporting the core description of the approach. It also describes earlier ancestral reconstructions of viral proteins, so this paper does not establish that such an approach was first reported in 2015. The passages have no page numbers in our copy. (LLM, conf=0.85)

  Experimental — accuracy not yet measured

May combine two claims.

Gradinaru's lab developed the CREATE platform and identified PhP.B and PhP.eB, which showed overwhelming superiority of CNS transduction after systemic delivery in some mouse strains but failed to translate into NHPs.

“Based on such strategy, Gradinaru’s lab developed Cre recombination-based AAV targeted evolution (CREATE) platform and successfully identified PhP.B and PhP.eB [64, 65], showing overwhelming superiority of CNS transduction after systemic delivery in some mouse strains but unfortunately failed to translate into NHPs [66, 67].”

• cites [64] Deverman BE, Pravdo PL, Simpson BP, Kumar SR, Chan KY, Banerjee A, Wu WL, Yang B, Huber N, Pasca SP, et al. Cre-dependent selection yields AAV variants for widespread gene transfer to the adult brain. Nat Biotechnol. 2016;34(2):204–9. sourcePartly supported

  The paper’s Abstract describes CREATE and identifies AAV-PHP.B, reporting at least 40-fold greater CNS gene transfer than AAV9 after intravenous injection in adult mice. It does not identify AAV-PHP.eB; the citing sentence pairs this paper with another citation for the variants. The NHP translation claim is attached to separate citations [66, 67], not to this source. (LLM, conf=0.87)

  Experimental — accuracy not yet measured
• cites [65] Chan KY, Jang MJ, Yoo BB, Greenbaum A, Ravi N, Wu WL, Sanchez-Guardado L, Lois C, Mazmanian SK, Deverman BE, et al. Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems. Nat Neurosci. 2017;20(8):1172–9. sourceSupported
• cites [66] Hordeaux J, Wang Q, Katz N, Buza EL, Bell P, Wilson JM. The neurotropic properties of AAV-PHPB are limited to C57BL/6J mice. Mol Ther. 2018;26(3):664–8. sourcePartly supported

  The Abstract and Main Text report that intravenous PHP.B produced much higher CNS transduction in C57BL/6J mice than AAV9, but that this advantage was not found in BALB/cJ mice or nonhuman primates. The source does not establish the same mouse-strain and NHP findings for PHP.eB. The CREATE-platform identification is attached to other citations in the citing sentence. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [67] Matsuzaki Y, Konno A, Mochizuki R, Shinohara Y, Nitta K, Okada Y, Hirai H. Intravenous administration of the adeno-associated virus-PHP.B capsid fails to upregulate transduction efficiency in the marmoset brain. Neurosci Lett. 2018;665:182–8. sourceNot checked — no full text of the source

May combine two claims.

Later, they optimized the CREATE platform by introducing a peptide library into the surface-exposed AA455 loop of PhP.eB capsid and obtained AAV.CAP-B10 with enhanced CNS tropism and simultaneously de-targeted from liver expression in both mice and marmosets.

“Later, they optimized the CREATE platform by introducing peptide library into the surface-exposed AA455 loop of PhP.eB capsid and obtained a variant AAV.CAP-B10 with enhanced CNS tropism and simultaneously de-targeted from liver expression in both mice and marmosets [68].”

• cites [68] Goertsen D, Flytzanis NC, Goeden N, Chuapoco MR, Cummins A, Chen Y, Fan Y, Zhang Q, Sharma J, Duan Y, et al. AAV capsid variants with brain-wide transgene expression and decreased liver targeting after intravenous delivery in mouse and marmoset. Nat Neurosci. 2022;25(1):106–15. sourceSupported

May combine two claims.

Incorporation of 5 copies of miR-122 target sequences has shown enhanced inhibitory effects over 3 copies in the liver of ICR mice, while without any significant change in transduction efficiency in 293 T cells.

“Incorporation of 5 copies of miR-122 target sequences has shown enhanced inhibitory effects over 3 copies in the liver of ICR mice, while without any significant change in transduction efficiency in 293 T cells [33].”

• cites [33] Qiao C, Yuan Z, Li J, He B, Zheng H, Mayer C, Li J, Xiao X. Liver-specific microRNA-122 target sequences incorporated in AAV vectors efficiently inhibits transgene expression in the liver. Gene Ther. 2011;18(4):403–10. sourcePartly supported

  The Results report greater reductions in liver luciferase expression with five miR-122 target copies than with three after plasmid injection; the Methods identify the mice used for hydrodynamic injection as ICR mice (pages unavailable). The Results also report no difference in luciferase activity in transfected 293 cells, but the paper does not identify them as 293T cells or measure transduction efficiency in that comparison. (LLM, conf=0.88)

  Experimental — accuracy not yet measured

May combine two claims.

Compared to systemic delivery, the intra-CSF route performs a more invasive procedure, but attenuates exposure to pre-existing neutralizing antibodies from the vascular system and avoids the risk of liver toxicity after intravenous administration of high doses.

“While comparing to systemic delivery, intra-CSF route performs in a more invasive procedure, but on the other hand, attenuates the exposure to pre-existing neutralizing antibodies from vascular system and avoids the risk of liver toxicity after intravenous administration of high doses [87].”

• cites [87] Lu CT, Zhao YZ, Wong HL, Cai J, Peng L, Tian XQ. Current approaches to enhance CNS delivery of drugs across the brain barriers. Int J Nanomedicine. 2014;9:2241–57. sourcePartly supported

  Split into 2 atoms — 1 confirmed, 1 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

  Experimental — accuracy not yet measured

May combine two claims.

AAV7 induces a stronger anti-AAV immune response in the hemophilia B population than AAV8, which makes AAV8 suitable for hemophilia B patients in clinical trials.

“Notably, AAV7 induces stronger anti-AAV immune response in the hemophilia B population than AAV8 [49], which makes AAV8 suitable for hemophilia B patients in clinical trials (Identifier: NCT01620801 and NCT00979238).”

• cites [49] Shao W, Pei X, Cui C, Askew C, Dobbins A, Chen X, Abajas YL, Gerber DA, Samulski RJ, Nichols TC, et al. Superior human hepatocyte transduction with adeno-associated virus vector serotype 7. Gene Ther. 2019;26(12):504–14. sourcePartly supported

  The Results section (page unavailable) reports neutralizing antibodies to AAV7 in all 26 hemophilia patients tested, versus no detectable antibodies to AAV8 in six. But the study measured pre-existing antibodies, not whether AAV7 induces a stronger immune response, and it does not conclude that this comparison makes AAV8 suitable for clinical trials. (LLM, conf=0.87)

  Experimental — accuracy not yet measured
• cites NCT01620801Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

• cites NCT00979238Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

May combine two claims.

AAV9 binds to terminal galactose and shows efficient transduction in CNS, eye, cardiac and skeletal muscle, and liver.

“Another validated vector with broad tropism is AAV9, binding to terminal galactose and showing efficient transduction in CNS, eye, cardiac and skeletal muscle, and liver [50–53].”

• cites [50] Gray SJ, Matagne V, Bachaboina L, Yadav S, Ojeda SR, Samulski RJ. Preclinical differences of intravascular AAV9 delivery to neurons and glia: a comparative study of adult mice and nonhuman primates. Mol Ther. 2011;19(6):1058–69. sourceNot checked — no full text of the source
• cites [51] Bell CL, Vandenberghe LH, Bell P, Limberis MP, Gao GP, Van Vliet K, Agbandje-McKenna M, Wilson JM. The AAV9 receptor and its modification to improve in vivo lung gene transfer in mice. J Clin Invest. 2011;121(6):2427–35. sourcePartly supported

  The Discussion identifies terminal β-galactose linkages as mediating AAV9 binding, and the Results report efficient liver transduction and efficient cardiac and skeletal muscle transduction at higher doses. The Results describe CNS transduction after intravenous injection as detectable, and the Discussion calls neuronal transduction limited—not efficient. The passages do not establish efficient eye transduction by AAV9. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [52] Fechner H, Sipo I, Westermann D, Pinkert S, Wang X, Suckau L, Kurreck J, Zeichhardt H, Muller O, Vetter R, et al. Cardiac-targeted RNA interference mediated by an AAV9 vector improves cardiac function in coxsackievirus B3 cardiomyopathy. J Mol Med (Berl). 2008;86(9):987–97. sourceNot checked — no full text of the source
• cites [53] Pacak CA, Mah CS, Thattaliyath BD, Conlon TJ, Lewis MA, Cloutier DE, Zolotukhin I, Tarantal AF, Byrne BJ. Recombinant adeno-associated virus serotype 9 leads to preferential cardiac transduction in vivo. Circ Res. 2006;99(4):e3-9. sourceNot checked — no full text of the source

May combine two claims.

Later studies characterized Anc80L65, demonstrating great transduction efficiency for retinas, inner ear, and notable expression in the CNS after systemic gene transfer.

“Later, more studies further characterized the vector Anc80L65, demonstrating great transduction efficiency for retinas [71], inner ear [72], and even notable expression in the CNS after systemic gene transfer [73].”

• cites [71] Carvalho LS, Xiao R, Wassmer SJ, Langsdorf A, Zinn E, Pacouret S, Shah S, Comander JI, Kim LA, Lim L, et al. Synthetic adeno-associated viral vector efficiently targets mouse and nonhuman primate retina in vivo. Hum Gene Ther. 2018;29(7):771–84. sourceSupported
• cites [72] Landegger LD, Pan B, Askew C, Wassmer SJ, Gluck SD, Galvin A, Taylor R, Forge A, Stankovic KM, Holt JR, et al. A synthetic AAV vector enables safe and efficient gene transfer to the mammalian inner ear. Nat Biotechnol. 2017;35(3):280–4. sourceSupported
• cites [73] Hudry E, Andres-Mateos E, Lerner EP, Volak A, Cohen O, Hyman BT, Maguire CA, Vandenberghe LH. Efficient gene transfer to the central nervous system by single-stranded Anc80L65. Mol Ther Methods Clin Dev. 2018;10:197–209. sourceSupported

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

Addition of SV40 intron confers about twofold enhancement of AAV transgene expression in lung carcinoma cells.

“Moreover, addition of SV40 intron confers about twofold enhancement of AAV transgene expression in lung carcinoma cells [25].”

• cites [25] Ostedgaard LS, Rokhlina T, Karp PH, Lashmit P, Afione S, Schmidt M, Zabner J, Stinski MF, Chiorini JA, Welsh MJ. A shortened adeno-associated virus expression cassette for CFTR gene transfer to cystic fibrosis airway epithelia. Proc Natl Acad Sci USA. 2005;102(8):2952–7. sourcePartly supported

  The Results and Discussion report increased β-gal activity with an SV40 intron in a full-length promoter construct and with the 113CMV promoter, but no increase with the 173CMV promoter. The paper does not state an approximately twofold increase, and it says the intron was not included in the AAV expression cassette. Materials and Methods describe the intron tests as plasmid-transfection experiments in cell lines, rather than tests of intron-containing AAV vectors. (LLM, conf=0.88)

  Experimental — accuracy not yet measured

May combine two claims.

Compared to IT route, ICV administration accesses the ventricles requiring an invasive stereotaxic surgery but has been demonstrated to be safe in clinical trials and achieve widespread AAV distribution in the brain.

“Comparing to IT route, ICV administration accesses the ventricles requiring an invasive stereotaxic surgery but has been demonstrated to be safe in clinical trials and achieve widespread AAV distribution in the brain [93].”

• cites [93] Duma C, Kopyov O, Kopyov A, Berman M, Lander E, Elam M, Arata M, Weiland D, Cannell R, Caraway C, et al. Human intracerebroventricular (ICV) injection of autologous, non-engineered, adipose-derived stromal vascular fraction (ADSVF) for neurodegenerative disorders: results of a 3-year phase 1 study of 113 injections in 31 patients. Mol Biol Rep. 2019;46(5):5257–72. sourcePartly supported

  The Ommaya reservoir implantation technique describes image-guided surgery to place a ventricular catheter, and the Safety section reports on ICV injections in a phase 1 trial. But the trial administered ADSVF cells, not AAV, and the paper does not demonstrate widespread AAV distribution in the brain. It also notes that patients with an existing VP shunt did not require reservoir implantation. (LLM, conf=0.90)

  Experimental — accuracy not yet measured

May combine two claims.

AAV-mediated gene therapy using IM delivery is minimally invasive in local muscle tissue and undergoes a cascade intracellular event for retrograde axonal trafficking into neuronal soma.

“AAV-mediated gene therapy using IM delivery is minimally invasive in local muscle tissue and undergoes a cascade intracellular event for retrograde axonal trafficking into neuronal soma [99, 100].”

• cites [99] Tosolini AP, Sleigh JN. Intramuscular delivery of gene therapy for targeting the nervous system. Front Mol Neurosci. 2020;13:129. sourceSupported
• cites [100] Chen Z, Fan G, Li A, Yuan J, Xu T. rAAV2-retro enables extensive and high-efficient transduction of lower motor neurons following intramuscular injection. Mol Ther Methods Clin Dev. 2020;17:21–33. sourcePartly supported

  The Discussion (page unavailable) describes intramuscular administration as minimally invasive and says rAAV can use axonal transport to travel toward lower motor neurons. It does not describe a cascade of intracellular events leading to the neuronal soma; it also reports cerebrospinal-fluid spread as a pathway for the extensive transduction observed in this study. (LLM, conf=0.83)

  Experimental — accuracy not yet measured

May combine two claims.

A novel variant AAV-TT derived from AAV2 revealed strong neurotropism and exceeded transduction of AAV9 and AAVrh10 in mice, even correcting neurological deficits in the mouse model of MPSIIIC.

“It is noted a novel variant AAV-TT derived from AAV2 revealed strong neurotropism and exceeded transduction of AAV9 and AAVrh10 in mice, even correcting neurological deficits in the mouse model of MPSIIIC [61].”

• cites [61] Tordo J, O’Leary C, Antunes A, Palomar N, Aldrin-Kirk P, Basche M, Bennett A, D’Souza Z, Gleitz H, Godwin A, et al. A novel adeno-associated virus capsid with enhanced neurotropism corrects a lysosomal transmembrane enzyme deficiency. Brain. 2018;141(7):2014–31. sourceSupported

May combine two claims.

Albright and colleagues identified essential 8 amino acid residues on the AAVrh.10 capsid that mediated transport across the BBB and grafted them onto AAV1 capsid, enabling the new chimeric capsid to cross the BBB and preferentially transduce neurons in C57/Bl6 mice.

“Moreover, Albright and colleagues identified the essential 8 amino acid residues on the AAVrh.10 capsid that mediated transport across the BBB and grafted onto AAV1 capsid, enabling the new chimeric capsid to cross the BBB and preferentially transduced neurons in C57/Bl6 mice [58].”

• cites [58] Albright BH, Storey CM, Murlidharan G, Castellanos Rivera RM, Berry GE, Madigan VJ, Asokan A. Mapping the structural determinants required for AAVrh.10 transport across the blood-brain barrier. Mol Ther. 2018;26(2):510–23. sourceSupported

May combine two claims.

Ramirez and colleagues demonstrated that AAV9 preferably crossed brain-endothelial barriers through an active, cell-mediated process, while little transduced endothelial cells.

“Ramirez and colleagues [115] demonstrated that AAV9 preferably crossed brain-endothelial barriers through an active, cell-mediated process, while little transduced endothelial cells.”

• cites [115] Merkel SF, Andrews AM, Lutton EM, Mu D, Hudry E, Hyman BT, Maguire CA, Ramirez SH. Trafficking of adeno-associated virus vectors across a model of the blood-brain barrier; a comparative study of transcytosis and transduction using primary human brain endothelial cells. J Neurochem. 2017;140(2):216–30. sourceSupported

May combine two claims.

AAV-mediated expression of AADC increases dopamine production and confers benefits to some PD patients with moderate motor improvement.

“Increasing dopamine production by AAV-mediated expression of the L-aromatic acid decarboxylase (AADC) enzyme, conferring benefits to some PD patients with moderately motor improvement [161]”

• cites [161] Christine CW, Richardson RM, Van Laar AD, Thompson ME, Fine EM, Khwaja OS, Li C, Liang GS, Meier A, Roberts EW, et al. Safety of AADC gene therapy for moderately advanced Parkinson disease: three-year outcomes from the PD-1101 trial. Neurology. 2022;98(1):e40–50. sourcePartly supported

  The introduction describes VY-AADC01 as designed to increase dopamine production, but says localized production is expected rather than reporting a direct measurement of increased dopamine. The Results report increased AADC activity and improved motor scores in participants with moderately advanced PD; the Discussion characterizes clinical benefit as potential. The claim overstates the certainty of increased dopamine production and benefit. (LLM, conf=0.85)

  Experimental — accuracy not yet measured

May combine two claims.

Optimal transduction of AAV2 vectors in the brain was achieved by IV injection of AAV 8 min after mannitol administration; delivery less than 5 min or 10 min after mannitol infusion was ten times less efficient.

“a study showed that the optimal transduction of AAV2 vectors in the brain was achieved by IV injection of AAV 8 min after mannitol administration; delivery of the vector less than 5 min or 10 min after mannitol infusion was ten times less efficient.”

• cites [133] McCarty DM, DiRosario J, Gulaid K, Muenzer J, Fu H. Mannitol-facilitated CNS entry of rAAV2 vector significantly delayed the neurological disease progression in MPS IIIB mice. Gene Ther. 2009;16(11):1340–52. sourcePartly supported

  Split into 2 atoms — 0 confirmed, 2 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

  Experimental — accuracy not yet measured

May combine two claims.

THR peptide increased AAV8 transcytosis in hCMEC/D3 endothelial cells and delivered more virion across the BBB to the brain parenchyma.

“further mechanistic study revealed the THR peptide increased the AAV8 transcytosis in hCMEC/D3 endothelial cells and delivered more virion across the BBB to the brain parenchyma [143].”

• cites [143] Zhang X, He T, Chai Z, Samulski RJ, Li C. Blood-brain barrier shuttle peptides enhance AAV transduction in the brain after systemic administration. Biomaterials. 2018;176:71–83. sourceSupported

May combine two claims.

AAVrh.8 better transduces neurons, astrocytes, and oligodendrocytes and has low peripheral tropism compared to AAV9 and AAVrh.10.

“AAVrh.8 was shown to better transduce neurons, astrocytes, and oligodendrocytes, while with low peripheral tropism compared to AAV9 and AAVrh.10 [126, 127].”

• cites [126] Tanguy Y, Biferi MG, Besse A, Astord S, Cohen-Tannoudji M, Marais T, Barkats M. Systemic AAVrh10 provides higher transgene expression than AAV9 in the brain and the spinal cord of neonatal mice. Front Mol Neurosci. 2015;8:36. sourceCould not verify

  The Materials and methods section describes vectors of serotypes AAV9 and AAVrh10, not AAVrh.8. The Results compare those two serotypes in the peripheral nervous system and in the heart and liver, but provide no AAVrh.8 comparison. A search for AAVrh.8/AAVrh8 found no passage addressing its cell transduction or peripheral tropism. (LLM, conf=0.77)

  Experimental — accuracy not yet measured
• cites [127] Zhang H, Yang B, Mu X, Ahmed SS, Su Q, He R, Wang H, Mueller C, Sena-Esteves M, Brown R, et al. Several rAAV vectors efficiently cross the blood-brain barrier and transduce neurons and astrocytes in the neonatal mouse central nervous system. Mol Ther. 2011;19(8):1440–8. sourceCould not verify

  The Discussion (page unavailable) identifies rAAVrh.10, rAAVrh.39, rAAVrh.43, and rAAV9 as effective CNS vectors, but does not assess AAVrh.8. Searches for AAVrh.8, oligodendrocytes, and peripheral tropism found no passage supporting the claimed comparison. (LLM, conf=0.76)

  Experimental — accuracy not yet measured

May combine two claims.

LY6A is responsible for specific engagement to the 7-mer insertion of PHP.B and PHP.eB and for transcytosis at the BBB.

“Later, it was reported that lymphocyte antigen 6 complex, locus A (LY6A), a GPI-anchored membrane protein, responsible for specific engagement to the 7-mer insertion of PHP.B and PHP.eB and transcytosis at BBB [118, 119].”

• cites [118] Huang Q, Chan KY, Tobey IG, Chan YA, Poterba T, Boutros CL, Balazs AB, Daneman R, Bloom JM, Seed C, et al. Delivering genes across the blood-brain barrier: LY6A, a novel cellular receptor for AAV-PHP.B capsids. PLoS One. 2019;14(11): e0225206. sourcePartly supported

  The Results section, “Interaction with Ly6a is necessary for the enhanced transduction phenotype of AAV-PHP.eB” (page unavailable), supports LY6A-dependent binding and transduction in mouse brain endothelial cells. The Discussion (page unavailable) reports increased binding by PHP.B variants with distinct seven-amino-acid insertions, but only suggests that LY6A could facilitate transcytosis across the C57BL/6J blood–brain barrier; it does not establish that transcytosis occurs through LY6A or that the insertion itself is the binding site. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [119] Hordeaux J, Yuan Y, Clark PM, Wang Q, Martino RA, Sims JJ, Bell P, Raymond A, Stanford WL, Wilson JM. The GPI-linked protein LY6A drives AAV-PHP.B Transport across the blood-brain barrier. Mol Ther. 2019;27(5):912–21. sourceNot checked — no full text of the source

May combine two claims.

AAV9-PB5-3 complex used to express IDUA in the MPS I mouse model achieved more than twofold higher expression with successful phenotypic correction in the brains.

“Thus, AAV9-PB5-3 complex was used to express IDUA in the mucopolysaccharidosis type I (MPS I) mouse model and achieved more than twofold higher expression with successful phenotypic correction in the brains.”

• cites [144] Zhang X, Chai Z, Lee Dobbins A, Itano MS, Askew C, Miao Z, Niu H, Samulski RJ, Li C. Customized blood-brain barrier shuttle peptide to increase AAV9 vector crossing the BBB and augment transduction in the brain. Biomaterials. 2022;281: 121340. sourcePartly supported

  The Results section on the IDUA−/− mouse model reports more than twofold higher brain IDUA activity with AAV9-PB5-3 than with AAV9 alone, and significantly lower GAG levels. The Discussion describes increased phenotypic correction in the brains of MPS I mice. The reported twofold measure was enzyme activity, however, not a direct measure of IDUA expression. (LLM, conf=0.84)

  Experimental — accuracy not yet measured

May combine two claims.

THR peptide can increase BBB-crossing efficiency of AAV9 and improve brain transduction in a dose-dependent manner in mice.

“can increase the BBB-crossing efficiency of AAV9 and improve brain transduction in a dose-dependent manner in mice.”

• cites [143] Zhang X, He T, Chai Z, Samulski RJ, Li C. Blood-brain barrier shuttle peptides enhance AAV transduction in the brain after systemic administration. Biomaterials. 2018;176:71–83. sourcePartly supported

  Results §3.2 (page unavailable) reports dose-dependent improvement in brain transduction in mice given THR-treated AAV8. Results §3.3 links THR to increased AAV8 crossing of the BBB. The cited experiments concern AAV8, not the AAV9 specified in the claim. (LLM, conf=0.90)

  Experimental — accuracy not yet measured

May combine two claims.

AAV-F achieved 65-fold higher astrocyte transduction and 171-fold higher neuron transduction than parental AAV9.

“with 65-fold and 171-fold higher transduction of astrocytes and neurons than parental AAV9 respectively [129].”

• cites [129] Hanlon KS, Meltzer JC, Buzhdygan T, Cheng MJ, Sena-Esteves M, Bennett RE, Sullivan TP, Razmpour R, Gong Y, Ng C, et al. Selection of an efficient AAV vector for robust CNS transgene expression. Mol Ther Methods Clin Dev. 2019;15:320–32. sourceSupported

May combine two claims.

AAV-PHP.B and AAV-PHP.eB show significantly higher efficiency in transporting across the BBB in C57BL/6J mice after intravenous delivery but are starkly absent in BALB/cJ mice and marmosets.

“Gradinaru lab identified AAV-PHP⋅B and AAV-PHP.eB with significant higher efficiencies in transporting across the BBB in C57BL/6 J mice after intravenous delivery but starkly absent in BALB/cJ mice and marmosets [64–67].”

• cites [64] Deverman BE, Pravdo PL, Simpson BP, Kumar SR, Chan KY, Banerjee A, Wu WL, Yang B, Huber N, Pasca SP, et al. Cre-dependent selection yields AAV variants for widespread gene transfer to the adult brain. Nat Biotechnol. 2016;34(2):204–9. sourcePartly supported

  The gene-transfer results (page unavailable) report significantly greater CNS gene transfer by intravenously delivered AAV-PHP.B than by AAV9, and the Vector biodistribution Methods specify C57Bl/6 mice. Our copy does not report AAV-PHP.eB or test whether either variant lacks this effect in BALB/cJ mice or marmosets. It also discusses BBB transport as a possible intermediate step rather than directly measuring greater BBB crossing. (LLM, conf=0.89)

  Experimental — accuracy not yet measured
• cites [65] Chan KY, Jang MJ, Yoo BB, Greenbaum A, Ravi N, Wu WL, Sanchez-Guardado L, Lois C, Mazmanian SK, Deverman BE, et al. Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems. Nat Neurosci. 2017;20(8):1172–9. sourcePartly supported

  The Discussion and Results report that AAV-PHP.B was selected for BBB crossing and that intravenously delivered AAV-PHP.eB efficiently transduced mouse brain cells. Methods (Animals; page unavailable) identifies C57BL/6J mice, but the paper does not report the claimed absence of BBB transport in BALB/cJ mice or marmosets. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [66] Hordeaux J, Wang Q, Katz N, Buza EL, Bell P, Wilson JM. The neurotropic properties of AAV-PHPB are limited to C57BL/6J mice. Mol Ther. 2018;26(3):664–8. sourcePartly supported

  The Abstract reports that intravenous AAV-PHP.B produced much higher CNS transduction in C57BL/6J mice than AAV9, but not in BALB/cJ mice. It also reports low CNS transduction in rhesus macaques, not marmosets. The cited paper does not establish the claim about AAV-PHP.eB. (LLM, conf=0.90)

  Experimental — accuracy not yet measured
• cites [67] Matsuzaki Y, Konno A, Mochizuki R, Shinohara Y, Nitta K, Okada Y, Hirai H. Intravenous administration of the adeno-associated virus-PHP.B capsid fails to upregulate transduction efficiency in the marmoset brain. Neurosci Lett. 2018;665:182–8. sourceNot checked — no full text of the source

May combine two claims.

PB5-3 peptide specifically binds to AAV9 serotype and enhances transduction efficiency in different brain cells in mice.

“Another PB5-3 peptide specifically bound to AAV9 serotype and enhanced transduction efficiency in different brain cells in mice [144].”

• cites [144] Zhang X, Chai Z, Lee Dobbins A, Itano MS, Askew C, Miao Z, Niu H, Samulski RJ, Li C. Customized blood-brain barrier shuttle peptide to increase AAV9 vector crossing the BBB and augment transduction in the brain. Biomaterials. 2022;281: 121340. sourceSupported

May combine two claims.

Ubiquitous promoters are frequently chosen to reach potent expression, with over 50% usage reported in 106 clinical trials and 76% in CNS disorders.

“Ubiquitous promoters are frequently chosen to reach potent expression, with over 50% usage reported in 106 clinical trials and 76% in CNS disorders.”

Classified as probably not a substantial claim.

Continuing efforts are being made to optimize rAAV vectors from their core genetic payloads to their capsid structure.

“Therefore, continuing efforts are being made to optimize the rAAV vectors from their “core” genetic payloads to their “coat” or capsid structure.”

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

Enhancer elements amplify transcriptional signals through physical contact with target promoters in the presence of RNA polymerase II, while located either upstream or downstream of the promoter.

“Enhancer element functions by amplifying transcriptional signals through physical contact with its target promoters in the presence of RNA polymerase II, while located either in the upstream or downstream of the promoter [19].”

• cites [19] Blackwood EM, Kadonaga JT. Going the distance: a current view of enhancer action. Science. 1998;281(5373):60–3. sourceNot checked — no full text of the source

May combine two claims.

Major challenges remain in applying rAAVs for CNS gene therapy, including suboptimal vector design, low CNS transduction efficiency and specificity, and therapy-induced immunotoxicity.

“However, major challenges remain in the application of rAAVs for CNS gene therapy, such as suboptimal vector design, low CNS transduction efficiency and specificity, and therapy-induced immunotoxicity.”

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

Recombinant adeno-associated viruses (rAAVs) are an attractive tool for gene delivery and have shown promise in gene therapy and gene editing, with potential one-and-done treatment benefits compared to conventional drugs.

“Recombinant adeno-associated viruses (rAAVs) have emerged as an attractive tool for gene delivery, and demonstrated tremendous promise in gene therapy and gene editing—therapeutic modalities with potential “one-and-done” treatment benefits compared to conventional drugs.”

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

AAV comprises a nonenveloped icosahedral viral capsid of approximately 26 nm in diameter, which encapsulates a linear single-stranded DNA genome of approximately 4.7 kb.

“AAV comprises of a nonenveloped icosahedral viral capsid of ~ 26 nm in diameter, which encapsulates a linear single-stranded DNA genome of ~ 4.7 kb [4].”

• cites [4] Tratschin JD, West MH, Sandbank T, Carter BJ. A human parvovirus, adeno-associated virus, as a eucaryotic vector: transient expression and encapsidation of the procaryotic gene for chloramphenicol acetyltransferase. Mol Cell Biol. 1984;4(10):2072–81. sourceNot checked — no full text of the source

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

AAV-mediated gene therapy has been galvanized by the marketing approval of Luxturna for RPE65-associated retinal dystrophy and Zolgensma for SMN1-linked spinal muscular atrophy.

“Recently, AAV-mediated gene therapy has been galvanized by the marketing approval of Luxturna for RPE65-associated retinal dystrophy and Zolgensma for SMN1-linked spinal muscular atrophy.”

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

The AAV genome contains symmetrical ITRs at each end, serving as the origin of DNA replication and packaging signals.

“The AAV genome contains symmetrical ITRs at each end, serving as the origin of DNA replication and packing signals [9].”

• cites [9] Wang XS, Ponnazhagan S, Srivastava A. Rescue and replication signals of the adeno-associated virus 2 genome. J Mol Biol. 1995;250(5):573–80. sourceNot checked — no full text of the source

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

More efforts are required in designing AAV vectors and optimizing therapeutic strategy in viral genome accommodation/optimization, capsid selection, administration route selection, and understanding potential toxicities.

“Despite these notable successes, more efforts are required in designing AAV vectors and optimizing the therapeutic strategy in areas including: (1) Viral genome accommodation and optimization; (2) AAV capsid selection for ideal efficiency and specificity; (3) Need-driven proper selection of administration routes; and (4) Understanding mechanisms of potential toxicities.”

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

AAV-based therapies are actively being investigated in preclinical studies and clinical trials for other neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and Huntington's disease.

“In addition, AAV-based therapies are actively being investigated in preclinical studies and clinical trials for other neurodegenerative diseases such as Alzheimer’s disease (AD), Parkinson’s disease (PD), and Huntington's disease (HD) [8].”

• cites [8] Deverman BE, Ravina BM, Bankiewicz KS, Paul SM, Sah DWY. Gene therapy for neurological disorders: progress and prospects. Nat Rev Drug Discov. 2018;17(9):641–59. sourceNot checked — no full text of the source

May combine two claims.

Pan and colleagues generated a CpG-free ITR with equivalent biological potency of transgene expression but with an approximately threefold reduction in yield.

“Thus, Pan and colleagues adopted rational engineering method to generate CpG-free ITR with equivalent biological potency of transgene expression, but with an approximately threefold reduction in yield [16].”

• cites [16] Pan X, Yue Y, Boftsi M, Wasala LP, Tran NT, Zhang K, Pintel DJ, Tai PWL, Duan D. Rational engineering of a functional CpG-free ITR for AAV gene therapy. Gene Ther. 2022;29(6):333–45. sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S4 (based).

The rAAV genome controls transgene expression level, cell-type specificity, duration, and other important aspects of AAV therapeutics.

“The rAAV (recombinant adeno-associated virus) genome controls the transgene expression level, cell-type specificity, duration, and other important aspects of AAV therapeutics.”

May combine two claims.

ssAAV requires host-cell DNA polymerase machinery to produce the complementary strand for transduction, which is a major limitation that slows transgene expression.

“The single-stranded AAV (ssAAV) requires the host-cell DNA polymerase machinery to produce the complementary strand for transduction which represents a major limitation that slows down the transgene expression.”

Probably not a substantial claim: may not pass S4 (based).

rAAVs have tropisms for the central nervous system across various species including humans.

“Given their tropisms for the central nervous system (CNS) across various species including humans, rAAVs have been extensively investigated in both pre-clinical and clinical studies targeting neurodegenerative disease.”

May combine two claims.

The promoter serves as a critical regulatory element for protein expression in the rAAV vector genome and determines expression level, cell- or tissue-specificity, and spatial and temporal characteristics.

“The promoter serves as a critical regulatory element for protein expression in the rAAV vector genome, and determines the expression level, cell- or tissue-specificity, and spatial and temporal characteristics [17].”

• cites [17] Goverdhana S, Puntel M, Xiong W, Zirger JM, Barcia C, Curtin JF, Soffer EB, Mondkar S, King GD, Hu J, et al. Regulatable gene expression systems for gene therapy applications: progress and future challenges. Mol Ther. 2005;12(2):189–211. sourceNot checked — no full text of the source

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

AAV is autonomously replication-defective unless aided by co-infection with a helper virus such as adenovirus, herpesvirus, or vaccinia virus.

“AAV was then characterized as an autonomously replication-defective virus unless aided by co-infection of a helper virus, such as adenovirus, herpesvirus, or vaccinia virus [2, 3].”

• cites [2] Berns KI, Linden RM. The cryptic life style of adeno-associated virus. BioEssays. 1995;17(3):237–45. sourceNot checked — no full text of the source
• cites [3] Casto BC, Atchison RW, Hammon WM. Studies on the relationship between adeno-associated virus type I (AAV-1) and adenoviruses I. Replication of AAV-1 in certain cell cultures and its effect on helper adenovirus. Virology. 1967;32(1):52–9. sourceNot checked — no full text of the source

May combine two claims.

AAV9 and AAV.rh10 provided widespread expression patterns beyond the injection site with glia- and neuron-tropism respectively.

“On the contrary, AAV9 and AAV.rh10 provided widespread expression patterns beyond the injection site with glia- and neuron-tropism respectively [78, 79].”

• cites [78] Watakabe A, Ohtsuka M, Kinoshita M, Takaji M, Isa K, Mizukami H, Ozawa K, Isa T, Yamamori T. Comparative analyses of adeno-associated viral vector serotypes 1, 2, 5, 8 and 9 in marmoset, mouse and macaque cerebral cortex. Neurosci Res. 2015;93:144–57. sourceNot checked — no full text of the source
• cites [79] Sondhi D, Hackett NR, Peterson DA, Stratton J, Baad M, Travis KM, Wilson JM, Crystal RG. Enhanced survival of the LINCL mouse following CLN2 gene transfer using the rh.10 rhesus macaque-derived adeno-associated virus vector. Mol Ther. 2007;15(3):481–91. sourceNot checked — no full text of the source

May combine two claims.

The BBB protects the brain by blocking entry of molecules greater than 400 Da into the CNS, concomitantly with inhibition of systemic delivery of therapeutic drugs.

“It protects the brain by blocking entry of molecules greater than 400 Da into the CNS, concomitantly with inhibition of systemic delivery of therapeutic drugs.”

May combine two claims.

AAV2 is the most extensively investigated serotype by far, known for its wide tissue tropism, especially in efficiently transducing neurons, which expands its clinical applications to neurological disorders.

“The most extensively investigated serotype by far is AAV2, known for its wide tissue tropism, especially in efficiently transducing neurons, which expands its clinical applications to neurological disorders.”

May combine two claims.

The superiority of AAV.CPP.16 over AAV9 can translate from mice to non-human primates, showing 6- to 249-fold enhancement across 4 mouse strains and fivefold enhancement in non-human primates in CNS transduction after intravenous delivery.

“Strikingly, the superiority of AAV.CPP.16 over AAV9 can translate from mice to non-human primates, showing 6- to 249-fold enhancement across 4 mouse strains and fivefold enhancement in non-human primates (NHPs) in CNS transduction after intravenous delivery [57].”

• cites [57] Yao Y, Wang J, Liu Y, Qu Y, Wang K, Zhang Y, Chang Y, Yang Z, Wan J, Liu J, et al. Variants of the adeno-associated virus serotype 9 with enhanced penetration of the blood-brain barrier in rodents and primates. Nat Biomed Eng. 2022;6(11):1257–71. sourceNot checked — no full text of the source

May combine two claims.

IV delivery is a non-invasive method and could take advantage of the dense vascular network in the brain and spinal cord for more uniform vector distribution but would require vectors to circumvent the obstacle of the BBB.

“IV delivery is a non-invasive method and could take advantage of the dense vascular network in the brain and spinal cord for more uniform vector distribution but would require vectors to circumvent the obstacle of BBB [110].”

• cites [110] Saraiva J, Nobre RJ, Pereira de Almeida L. Gene therapy for the CNS using AAVs: The impact of systemic delivery by AAV9. J Control Release. 2016;241:94–109. sourceNot checked — no full text of the source

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

Since the discovery of AAV, 13 AAV serotypes and more than 150 variants have been naturally isolated from human, non-human primates, and other animal species.

“Since the discovery of AAV as contaminants from adenovirus stocks, 13 AAV serotypes and more than 150 variants have been naturally isolated from human, non-human primates, and other animal species.”

May combine two claims.

Direct CNS parenchymal injection is the most selected route for CNS diseases and can precisely transfer high concentrations of vectors to the target region via surgical approach, while bypassing the blockade of physiological barriers.

“The direct CNS parenchymal injection is the most selected route for CNS diseases and can precisely transfer high concentrations of vectors to the target region via surgical approach, while bypassing the blockade of physiological barriers.”

May combine two claims.

AAV-DJ can only transduce a confined region but with higher efficiency than AAV2, suggesting an ideal vector for desired expression within a small area.

“Similarly, AAV-DJ can only transduce a confined region but with higher efficiency than AAV2, suggesting an ideal vector for desired expression within a small area [63].”

• cites [63] Grimm D, Lee JS, Wang L, Desai T, Akache B, Storm TA, Kay MA. In vitro and in vivo gene therapy vector evolution via multispecies interbreeding and retargeting of adeno-associated viruses. J Virol. 2008;82(12):5887–911. sourceNot checked — no full text of the source

May combine two claims.

Successful gene therapies for neurological disorders rely on careful design of AAV vector, including optimization of genome cassette and selection of capsid, and require the most appropriate route for AAV administration.

“Successful gene therapies for neurological disorders not only rely on the careful design of AAV vector, including optimization of genome cassette and selection of capsid, but also require the most appropriate route for AAV administration (Fig. 2).”

May combine two claims.

AAV8 shows a broad range of tropism and has already been applied in clinical use for eye disorders and Lysosomal Storage Disorders.

“On the other hand, AAV8 shows a broad range of tropism and has already been applied in clinical use (Identifier: NCT03001310 and NCT04611503 for eye disorders, NCT04174105 and NCT03173521 for Lysosomal Storage Disorders).”

• cites NCT03001310Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

• cites NCT04611503Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

• cites NCT04174105Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

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

AAV2-based LUXTURNA® has been approved by FDA in 2017 for use in patients with Leber's congenital amaurosis.

“For instance, AAV2-based LUXTURNA® has been approved by FDA in 2017 for use in patients with Leber's congenital amaurosis [38].”

• cites [38] Russell S, Bennett J, Wellman JA, Chung DC, Yu ZF, Tillman A, Wittes J, Pappas J, Elci O, McCague S, et al. Efficacy and safety of voretigene neparvovec (AAV2-hRPE65v2) in patients with RPE65-mediated inherited retinal dystrophy: a randomised, controlled, open-label, phase 3 trial. Lancet. 2017;390(10097):849–60. sourceNot checked — no full text of the source

May combine two claims.

Receptor-mediated transcytosis (RMT) is the most extensively investigated non-invasive way to deliver AAV through the BBB into the CNS, although the choice of receptors responsible for AAV trafficking across the BBB is still not well understood.

“In a non-invasive way to deliver AAV through the BBB into the CNS, the receptor-mediated transcytosis (RMT) is the most extensively investigated, although it is still not well understood the choice of receptors responsible for AAV trafficking across the BBB.”

May combine two claims.

AAV9 holds the natural ability to cross the BBB and transduces the CNS after systemic administration.

“Moreover, AAV9 holds the natural ability to cross the BBB and transduces the CNS after systemic administration that will be further discussed in the sections below [54].”

• cites [54] Zincarelli C, Soltys S, Rengo G, Rabinowitz JE. Analysis of AAV serotypes 1–9 mediated gene expression and tropism in mice after systemic injection. Mol Ther. 2008;16(6):1073–80. sourceNot checked — no full text of the source

May combine two claims.

IT route injects therapeutic agents via lumbar puncture, which is the minimal invasive and easy access procedure for CSF delivery, transporting most molecules into the spinal cord with little transduction to the brain.

“IT route injects therapeutic agents via lumbar puncture, which is the minimal invasive and easy access procedure for CSF delivery, transporting most of molecules into the spinal cord with little transduction to the brain.”

May combine two claims.

AAV2 shows minimal diffusion and primarily transduces neurons, and has been the most extensively used vector for IP delivery of therapeutic genes in clinical trials for neurodegenerative diseases, such as Alzheimer's disease and Parkinson's disease.

“AAV2 shows minimal diffusion and primarily transduces neurons, which has the most extensively used vector to IP delivery of therapeutic genes in clinical trials for neurodegenerative diseases, such as Alzheimer’s disease (Identifier: NCT04133454 and NCT00876863) and Parkinson’s disease (Identifier: NCT04167540, NCT00643890 and NCT02418598).”

• cites NCT04133454Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

• cites NCT00876863Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

• cites NCT04167540Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based). May combine two claims.

AAV7 and AAV8 represent hepatocyte tropism, though their primary receptors are still unknown.

“Both, AAV7 and AAV8, represent hepatocyte tropism, though their primary receptors are still unknown.”

May combine two claims.

AAV1-driven release of single-chain variable fragment (scFv) antibodies targets Aβ deposition and attenuates cognitive deficits, preventing and treating Alzheimer's disease.

“AAV1-driven release of single-chain variable fragment (scFv) antibodies targets Aβ deposition and attenuates cognitive deficits, preventing and treating of Alzheimer’s disease [108, 109].”

• cites [108] Wang YJ, Gao CY, Yang M, Liu XH, Sun Y, Pollard A, Dong XY, Wu XB, Zhong JH, Zhou HD, et al. Intramuscular delivery of a single chain antibody gene prevents brain Abeta deposition and cognitive impairment in a mouse model of Alzheimer’s disease. Brain Behav Immun. 2010;24(8):1281–93. sourceNot checked — no full text of the source
• cites [109] Wang QH, Wang YR, Zhang T, Jiao SS, Liu YH, Zeng F, Li J, Yao XQ, Zhou HD, Zhou XF, et al. Intramuscular delivery of p75NTR ectodomain by an AAV vector attenuates cognitive deficits and Alzheimer’s disease-like pathologies in APP/PS1 transgenic mice. J Neurochem. 2016;138(1):163–73. sourceNot checked — no full text of the source

May combine two claims.

In the CNS, endogenous miR-124 can specifically de-target transduction of viral vectors in astrocytes in mouse brain, conferring the possibility to cell type-specific engineering.

“In the CNS, the endogenous miR-124 can specifically de-target transduction of viral vectors in astrocytes in mouse brain, conferring the possibility to cell type-specific engineering [34].”

• cites [34] Colin A, Faideau M, Dufour N, Auregan G, Hassig R, Andrieu T, Brouillet E, Hantraye P, Bonvento G, Deglon N. Engineered lentiviral vector targeting astrocytes in vivo. Glia. 2009;57(6):667–79. sourceNot checked — no full text of the source

May combine two claims.

Limitations of intra-CSF delivery include unfavorable flow dynamics of CSF and the huge size of human CNS organs often restricting virions to diffuse into deep CNS regions, and surgical procedures of ICM and ICV injection posing high-risk injury to the brain.

“However, the limitation of intra-CSF delivery still needs to be addressed: (1) unfavorable flow dynamics of CSF and the huge size of human CNS organs often restrict the virions to diffuse into the deep CNS regions; and (2) surgical procedure of ICM and ICV injection posing high-risk injury to the brain.”

May combine two claims.

AAV2 without BBB-crossing ability was found to be endocytosed to transduce BMVEC, hardly to diffuse across the endothelial barrier.

“In contrary, AAV2 without BBB-crossing ability was found to be endocytosed to transduce BMVEC, hardly to diffuse across the endothelial barrier.”

May combine two claims.

TJ loss is observed in the white matter of MS lesions and is positively correlated with BBB breakdown.

“TJ loss was also observed in the white matter of MS lesions and positively correlated with BBB breakdown [124].”

• cites [124] Kirk J, Plumb J, Mirakhur M, McQuaid S. Tight junctional abnormality in multiple sclerosis white matter affects all calibres of vessel and is associated with blood-brain barrier leakage and active demyelination. J Pathol. 2003;201(2):319–27. sourceNot checked — no full text of the source

May combine two claims.

Several clinical trials using IP-administered AAV2 gene therapies for AD have reported no positive outcomes despite demonstrating safety.

“Despite demonstrating safeties, several clinical trials using IP-administered AAV2 gene therapies so far have unfortunately reported no positive outcomes in treating AD [156].”

• cites [156] Rafii MS, Tuszynski MH, Thomas RG, Barba D, Brewer JB, Rissman RA, Siffert J, Aisen PS. Adeno-Associated Viral Vector (Serotype 2)-nerve growth factor for patients with alzheimer disease: a randomized clinical trial. JAMA Neurol. 2018;75(7):834–41. sourceNot checked — no full text of the source

May combine two claims.

Mannitol-mediated BBB disruption is temporary and reversible; BBB permeability peaks 5 min after intra-arterial infusion and lasts 20-30 min.

“As the mannitol-mediated BBB disruption is temporary and reversible, the BBB permeability peaks 5 min after intra-arterial (IA) infusion and lasts for 20–30 min [131, 132].”

• cites [131] Ikeda M, Bhattacharjee AK, Kondoh T, Nagashima T, Tamaki N. Synergistic effect of cold mannitol and Na(+)/Ca(2+) exchange blocker on blood-brain barrier opening. Biochem Biophys Res Commun. 2002;291(3):669–74. sourceNot checked — no full text of the source
• cites [132] Rapoport SI. Advances in osmotic opening of the blood-brain barrier to enhance CNS chemotherapy. Expert Opin Investig Drugs. 2001;10(10):1809–18. sourceNot checked — no full text of the source

May combine two claims.

AAVHSCs are neurotropic AAVs identified from healthy CD34+ human hematopoietic stem cells.

“AAVHSCs are another group of neurotropic AAVs identified from healthy CD34 + human hematopoietic stem cells [128].”

• cites [128] Smith LJ, Ul-Hasan T, Carvaines SK, Van Vliet K, Yang E, Wong KK Jr, Agbandje-McKenna M, Chatterjee S. Gene transfer properties and structural modeling of human stem cell-derived AAV. Mol Ther. 2014;22(9):1625–34. sourceNot checked — no full text of the source