Ionizable Lipid Nanoparticles for Therapeutic Base Editing of Congenital Brain Disease

May combine two claims.

The 12-LNP library had hydrodynamic diameters ranging from 67.8 to 128.3 nm with a maximum polydispersity value of 0.154, indicating small and relatively monodisperse LNPs.

“Hydrodynamic diameter for all LNP formulations – measured via dynamic light scattering intensity – ranged from 67.8 to 128.3 nm with a maximum polydispersity value of 0.154 (Fig. 1B), indicating small and relatively monodisperse LNPs (PDI < 0.3) (19).”

• cites [19] sourceDisputed

  The Results section, “Characterization of the LNP library” (Fig. 2C; page not provided), reports 14 formulations with DLS-measured diameters of 64.6–135.2 nm, not 12 formulations ranging from 67.8–128.3 nm. It also says one formulation had PDI above 0.3, contradicting the claimed maximum of 0.154, although the others were described as monodisperse. (LLM, conf=0.94)

  Checked by a reasoning model — may be wrong.

  Experimental — accuracy not yet measured

May combine two claims.

Viral delivery platforms can be limited by pre-existing viral immunity, transgene size constraints, undesired vector integration, and adverse events related to the viral vector.

“Although viral vectors are a promising approach to deliver gene editing technology to the CNS, viral delivery platforms can be limited by pre-existing viral immunity, transgene size constraints, undesired vector integration, and adverse events related to the viral vector (10–12).”

• cites [10] sourceSupported
• cites [11] sourceSupported
• cites [12] sourceNot checked — no full text of the source

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

Ionizable lipid nanoparticles (LNPs) are a promising non-viral delivery platform for mRNA nanomedicines and have been used to deliver siRNA and CRISPR-based gene editing therapies to the liver in postnatal mouse, non-human primate, and clinical studies with encouraging results.

“Ionizable lipid nanoparticles (LNPs) are a promising non-viral delivery platform for mRNA nanomedicines (13, 14) that have been used to deliver siRNA and CRISPR-based gene editing therapies to the liver in postnatal mouse, non-human primate, and clinical studies with encouraging results (15–18).”

• cites [13] sourceNot checked — no full text of the source
• cites [14] sourceNot checked — no full text of the source
• cites [15] 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
• cites [16] sourceNot checked — no full text of the source
• cites [17] sourceSupported
• cites [18] 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.

In utero gene editing has rescued the lethal phenotype of hereditary tyrosinemia type 1, improved the pulmonary phenotype of surfactant protein C deficiency, and ameliorated metabolic, musculoskeletal, and cardiac disease in mucopolysaccharidosis type I (MPS-IH, Hurler syndrome) in mouse models.

“Previously, we demonstrated the therapeutic potential of in utero gene editing in mouse models of human diseases, rescuing the lethal phenotype of hereditary tyrosinemia type 1, improving the pulmonary phenotype of surfactant protein C deficiency, and ameliorating metabolic, musculoskeletal, and cardiac disease in mucopolysaccharidosis type I (MPS-IH, Hurler syndrome) (7–9).”

• cites [7] sourcePartly supported

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

  Experimental — accuracy not yet measured
• cites [8] sourceSupported
• cites [9] sourceSupported

Classified as probably not a substantial claim.

Advances in mRNA-based gene editing tools, including CRISPR-Cas9 and base editing platforms, provide an opportunity for one-and-done treatments for monogenic diseases.

“Advances in mRNA-based gene editing tools, including CRISPR-Cas9 and base editing platforms, provide an opportunity for “one-and-done” treatments for monogenic diseases (5, 6).”

• cites [5] sourcePartly supported

  The Abstract describes CRISPR-Cas9 editing as a potential treatment for monogenic disorders, and the Delivery section discusses mRNA delivery of editing nucleases. Our copy does not discuss base-editing platforms or establish that treatment would require only one administration; those parts of the claim are unsupported. (LLM, conf=0.86)

  Experimental — accuracy not yet measured
• cites [6] sourceNot checked — no full text of the source

May combine two claims.

For many genetic CNS diseases, pathology begins before birth and is difficult to reverse after onset, resulting in significant morbidity by the time of or shortly after birth.

“Furthermore, the pathology of many genetic CNS diseases begins before birth and is difficult to reverse after onset resulting in significant morbidity by the time of or shortly after birth (3).”

• cites [3] sourcePartly supported

  The ‘In utero gene therapy: Rationale and supporting animal studies’ section says irreversible neurologic pathology begins before birth in neuronopathic Gaucher disease. The Conclusions also identify severe morbidity and irreversible early pathology as characteristics of potential target diseases. The paper does not establish that many genetic CNS diseases cause significant morbidity by birth or shortly afterward; it notes that a child can appear normal at birth despite fetal pathology. Page numbers were not available in our copy. (LLM, conf=0.84)

  Experimental — accuracy not yet measured

May combine two claims.

Similar to patients with MPS-IH, Idua-W392X mice have undetectable α-L-iduronidase (IDUA) enzyme activity and elevated tissue glycosaminoglycans (GAGs).

“Similar to patients with MPS-IH, Idua-W392X mice have undetectable α-L- iduronidase (IDUA) enzyme activity and elevated tissue glycosaminoglycans (GAGs) (32).”

• cites [32] sourceSupported

Classified as probably not a substantial claim.

The ideal N:P ratio is distinct for each ionizable lipid.

“The ideal N:P ratio is known to be distinct for each ionizable lipid (18).”

• cites [18] sourceCould not verify

  The Multi-tail ionizable lipids section discusses optimizing LNP composition, but does not identify an ideal N:P ratio or say that it differs for each ionizable lipid. Searches for N:P, nitrogen-to-phosphate ratio, and lipid-specific charge-ratio optimization found no passage addressing that point; page numbers were unavailable in our copy. (LLM, conf=0.74)

  Experimental — accuracy not yet measured

May combine two claims.

One of the most common IDUA disease-causing mutations is amenable to correction by adenine base editing resulting in restoration of IDUA enzyme activity and improvement in disease pathology including the reduction in lysosomal accumulation of GAGs.

“One of the most common IDUA disease-causing mutations is amenable to correction by adenine base editing resulting in restoration of IDUA enzyme activity and improvement in disease pathology including the reduction in lysosomal accumulation of GAGs (9).”

• cites [9] sourceSupported

Classified as probably not a substantial claim.

The molar ratios at which LNP components are combined can be optimized for specific applications to better encapsulate nucleic acid cargo, enhance uptake by target cells, or alter biodistribution and protein corona formation.

“The molar ratios at which the components of LNPs are combined can be optimized for specific applications to better encapsulate nucleic acid cargo, enhance uptake by target cells, or alter biodistribution and protein corona formation (28).”

• cites [28] sourcePartly supported

  The Introduction and Discussion state that LNP component ratios can be optimized for delivery and biodistribution, and that lipid-to-cargo ratios can improve encapsulation. The Results link excipient ratios to mRNA delivery, but do not measure target-cell uptake. The Discussion treats differences in protein corona formation as a hypothesis requiring future study, not an established effect of changing molar ratios. (LLM, conf=0.86)

  Experimental — accuracy not yet measured

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

Modulating excipient molar ratios improves in vitro delivery to fetal lung cells and corresponds to enhanced in vivo performance following intra-amniotic LNP injection.

“In prior work, we demonstrated that modulating excipient molar ratios improves in vitro delivery to fetal lung cells and corresponds to enhanced in vivo performance following intra-amniotic LNP injection (28).”

• cites [28] sourceSupported

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

The ionizable lipid is a major determinant of intracellular delivery by both directly complexing with mRNA and facilitating endosomal escape, while cholesterol provides stability and enables membrane fusion, DOPE supplies structural support, and PEG reduces aggregation and non-specific endocytosis.

“While the ionizable lipid – in this case C3 – is a major determinant of intracellular delivery by both directly complexing with mRNA and facilitating endosomal escape, the remaining excipients also play critical roles: cholesterol provides stability and enables membrane fusion, DOPE supplies structural support, and PEG reduces aggregation and non-specific endocytosis (29).”

• cites [29] 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.

The low-level editing observed via the current approach may beneficially affect the neurobehavioral phenotype in MPS-IH, although assessment of this outcome is limited in the current study due to the lack of a strong neurobehavioral phenotype in this mouse model at the short time points assessed.

“the low-level editing observed via the current approach may beneficially affect the neurobehavioral phenotype in MPS-IH, although assessment of this outcome is limited in the current study due to the lack of a strong neurobehavioral phenotype in this mouse model at the short time points assessed (9).”

• cites [9] 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.

A mouse model of MPS-IH was chosen to study LNP-mediated perinatal delivery of a gene editing therapy since the neurodevelopmental pathology in patients with this disease starts prior to or shortly after birth, is irreversible, and is resistant to currently available treatments, including enzyme replacement therapy and hematopoietic stem cell transplant.

“In this work, a mouse model of MPS-IH was chosen to study LNP-mediated perinatal delivery of a gene editing therapy since the neurodevelopmental pathology in patients with this disease starts prior to or shortly after birth, is irreversible, and is resistant to currently available treatments, including enzyme replacement therapy and hematopoietic stem cell transplant (9).”

• cites [9] 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.

The immune system of a P0 mouse is immature, similar to that of a developing human fetus, and a mature immune system, as is present in an adult mouse, is capable of producing an anti-PEG IgM response to C3.MPS LNPs.

“These data are consistent with our previous study demonstrating that the immune system of a P0 mouse is immature (35) – similar to that of a developing human fetus – and imply that a mature immune system, as is present in an adult mouse, is capable of producing an anti-PEG IgM response to C3.MPS LNPs.”

• cites [35] sourceSupported

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

The N:P ratio impacts both nucleic acid delivery and toxicity, with higher ratios leading to diminishing returns in cellular transfection due to increased cell death.

“Previous studies have demonstrated that the N:P ratio impacts both nucleic acid delivery and toxicity, with higher ratios leading to diminishing returns in cellular transfection due to increased cell death (31).”

• cites [31] sourcePartly supported

  The Introduction (page not provided) says high N:P ratios can enhance intracellular delivery and in vitro gene expression, with higher toxicity as a trade-off. It does not say that cell death causes diminishing returns in cellular transfection; the toxicity it describes is systemic and linked to interactions with blood components. (LLM, conf=0.88)

  Experimental — accuracy not yet measured

May combine two claims.

Since congenital brain pathology often begins before birth, fetal intervention provides a therapeutic opportunity to mitigate irreversible disease burden.

“Since congenital brain pathology often begins before birth, fetal intervention provides a therapeutic opportunity to mitigate irreversible disease burden (3).”

• cites [3] sourcePartly supported

  The ‘In utero gene therapy: Rationale and supporting animal studies’ section says irreversible neurologic pathology begins in utero in neuronopathic Gaucher disease and describes fetal gene therapy mitigating neuroinflammation and neurodegeneration in mice. This supports the therapeutic opportunity, but the paper does not establish that congenital brain pathology generally or often begins before birth. (LLM, conf=0.82)

  Experimental — accuracy not yet measured

Classified as probably not a substantial claim.

The molecular composition of an LNP modulates the endogenous proteins that bind the nanoparticle and govern its biological fate.

“Prior studies have demonstrated that the molecular composition of an LNP modulates the endogenous proteins that bind the nanoparticle and govern its biological fate (43).”

• cites [43] sourceSupported

May combine two claims.

In vivo, C3 LNPs formulated at 10:1 and 20:1 N:P ratios had similar performance in the neonatal brain to the initial screening study, with no significant difference between the LNPs formulated at different N:P ratios.

“IVIS imaging revealed that while both LNP formulations had similar performance in the neonatal brain to the initial screening study, there was no significant difference between the LNPs formulated at different N:P ratios (Fig. 3D).”

May combine two claims.

C3.GFP LNPs resulted in a high transfection efficiency with no significant effect on viability of neurons cultured from cerebral cortex tissue of two pediatric patients.

“C3.GFP LNPs resulted in a high transfection efficiency (Fig. 5F) with no significant effect on viability (Fig. 5G) of these cells.”

May combine two claims.

IVIS imaging of neonatal brains confirmed the performance trends observed in the fetal brain, with C3 LNPs facilitating 8-fold greater mRNA expression in the neonatal brain than the industry standard LNP (p < 0.0001).

“IVIS imaging of neonatal brains confirmed the performance trends observed in the fetal brain (Fig. 1F and 1G), with C3 LNPs facilitating 8-fold greater mRNA expression in the neonatal brain than the industry standard LNP (p < 0.0001).”

May combine two claims.

Each ionizable lipid produced LNPs with variable performance, demonstrating the crucial role of this component for mRNA delivery.

“Each ionizable lipid produced LNPs with variable performance, demonstrating the crucial role of this component for mRNA delivery (Fig. 1F).”

May combine two claims.

The zeta potential of C3.MPS LNPs did not change at increasing concentrations of human CSF, implying that these LNPs avoid significant protein adhesion that can reduce cellular uptake.

“However, the zeta potential of C3.MPS LNPs did not change at increasing concentrations of human CSF (Fig. S10C), implying that these LNPs avoid significant protein adhesion that can reduce cellular uptake.”

May combine two claims.

No editing above baseline was noted in genomic DNA isolated from the liver and gonads, demonstrating similar brain specific delivery observed in the initial LNP library screen.

“Notably, no editing above baseline was noted in genomic DNA isolated from the liver and gonads (Fig. S9), demonstrating similar brain specific delivery observed in our initial LNP library screen.”

May combine two claims.

In vivo transfection efficacy in the fetal brain showed trends with decreasing pKa, increasing size, and increasing zeta potential.

“Interestingly, we observed trends for in vivo transfection efficacy in the fetal brain with decreasing pKa, increasing size, and increasing zeta potential (Fig. S2).”

May combine two claims.

At one month, mice treated with C3.MPS LNPs demonstrated increased IDUA activity in the midbrain (p < 0.001, 6.97% of normal) and hippocampus (p < 0.0001, 11.3% of normal) relative to untreated Idua-W392X mice, associated with a reduction in GAG levels in the respective brain regions.

“At one month, mice treated with C3.MPS LNPs demonstrated increased IDUA activity in the midbrain (p < 0.001, 6.97% of normal) and hippocampus (p < 0.0001, 11.3% of normal) relative to untreated Idua-W392X mice that was associated with a reduction in GAG levels in the respective brain regions (Figs. 4C and 4D).”

May combine two claims.

Although whole brain distribution was similar across LNPs, the magnitude of GFP expression was proportional to LNP performance in initial screening studies (C3 > C2 > C1).

“Although we observed similarities in whole brain distribution across LNPs, the magnitude of GFP expression and was proportional to LNP performance in our initial screening studies (C3 > C2 > C1) (Fig. 2B).”

May combine two claims.

C3 LNPs encapsulating ABE7.10 mRNA and sgRNA demonstrated dose-dependent A→G editing of the target adenine in the mouse Idua gene and no toxicity in MPS-IH mouse fibroblasts or neurons.

“These studies demonstrated dose-dependent A→G editing of the target adenine in the mouse Idua gene (Fig. 3F) and no toxicity in either cell type (Fig. S8C).”

May combine two claims.

Despite low-level LNP-mediated adenine base editing at the pathogenic locus in the neonatal MPS-IH mouse brain, partial restoration of IDUA enzyme activity (7-11% of normal) and reduction in the accumulation of GAGs in multiple brain regions were observed.

“Despite low-level LNP-mediated adenine base editing at the pathogenic locus in the neonatal MPS-IH mouse brain, we observed partial restoration of IDUA enzyme activity (7–11% of normal) and reduction in the accumulation of GAGs in multiple brain regions.”

May combine two claims.

After ICV administration of C3 LNPs to E18 Balb/c fetuses, strong luciferase expression was observed in the fetal brain but no significant expression in the heart, lung, intestine, liver, spleen, or kidney.

“We observed strong luciferase expression in the fetal brain but no significant expression in the heart, lung, intestine, liver, spleen, or kidney (Fig. S4).”

May combine two claims.

In comparison to the original N:P ratio of 10:1, C3 LNPs formulated at an N:P ratio of 20:1 had a significant improvement in mRNA delivery (p < 0.001) albeit with enhanced toxicity.

“In comparison to the original N:P ratio of 10:1, LNPs formulated at an N:P ratio of 20:1 had a significant improvement in mRNA delivery (Fig. 3C, p < 0.001) albeit with enhanced toxicity (Fig. S8B).”

May combine two claims.

No significant change in size or polydispersity of C3.MPS LNPs incubated in human CSF was observed, and this result was confirmed over a one-week time course, supporting the stability of the LNP.

“No significant change in size or polydispersity of C3.MPS LNPs incubated in human CSF was observed (Fig. 5D and Fig. 5E), and this result was confirmed over a one-week time course, supporting the stability of our LNP.”

The quote only partly states this claim.

Modulation of LNP lipid ratio parameters suggested that the original LNP formulation mediated efficient mRNA delivery in neuronal origin cells.

“Modulation of LNP lipid ratio parameters suggested that our original LNP formulation mediated efficient mRNA delivery.”

May combine two claims.

The trends observed for in vivo transfection efficiency of the LNP library in the fetal mouse brain matched those seen in the neonatal mouse brain, validating that the neonatal mouse model can be used to approximate LNP performance in the fetal mouse CNS.

“More broadly, the trends observed for in vivo transfection efficiency of our LNP library in the fetal mouse brain matched those seen in the neonatal mouse brain, validating that the neonatal mouse model can be used to approximate LNP performance in the fetal mouse CNS.”

May combine two claims.

LNPs containing the C3 ionizable lipid were small (<100 nm), monodisperse (<0.20 PDI), well-encapsulated (>90%), and on the lower end of the known pKa range (~5) for efficient mRNA delivery.

“LNPs containing the C3 ionizable lipid were small (<100 nm), monodisperse (<0.20 PDI), well-encapsulated (>90%), and on the lower end of the known pKa range (~5) for efficient mRNA delivery.”

May combine two claims.

C3 LNPs delivered mRNA to the fetal NHP brain via ICV administration at a gestational age akin to a mid-gestation human fetus, resulting in transfection of the brain ventricular lining.

“To this end, we evaluated the ability of C3 LNPs to deliver mRNA to the fetal brain in the NHP via ICV administration at a gestational age akin to a mid-gestation human fetus and demonstrated transfection of the brain ventricular lining akin to our studies in the mouse model.”

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

Survival after neonatal ICV injection was 100% in R26mT/mG neonates.

“Survival after neonatal ICV injection was 100% in R26mT/mG neonates.”

May combine two claims.

This proof-of-concept study supports the safety and efficacy of LNPs for delivery of mRNA-based gene editing therapies to the CNS.

“This proof-of-concept study supports the safety and efficacy of LNPs for delivery of mRNA-based gene editing therapies to the CNS.”

Classified as probably not a substantial claim.

Through correction of underlying pathogenic mutations, gene editing technologies offer a potential curative strategy for genetic diseases with CNS features after a single treatment.

“Through correction of underlying pathogenic mutations, gene editing technologies offer a potential curative strategy for genetic diseases with CNS features after a single treatment.”

May combine two claims.

While C3.MPS LNPs produce an acute serum cytokine response that is generally pro-inflammatory, this reaction is at a lower level than that produced by FDA-approved MC3 LNPs.

“Taken together, these data suggest that while C3.MPS LNPs do produce an acute serum cytokine response that is generally pro-inflammatory, this reaction is at a lower level than that produced by FDA-approved MC3 LNPs.”

May combine two claims.

LNP encapsulation protects mRNA cargo from systemic degradation and facilitates intracellular delivery.

“All LNP formulations yielded greater mRNA delivery than injection of naked mRNA, confirming that LNP encapsulation protects mRNA cargo from systemic degradation and facilitates intracellular delivery (Fig. 1F).”

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

Non-human primate models have more structurally similar brain anatomy to humans and allow for the evaluation of the delivery approach, a minimally invasive, ultrasound-guided ICV, that would be used clinically.

“Relative to fetal and neonatal mouse models, non-human primate models have more structurally similar brain anatomy to humans and allow for the evaluation of the delivery approach, a minimally invasive, ultrasound-guided ICV, that would be used clinically.”

May combine two claims.

C3.MPS LNPs facilitate partial biochemical correction of disease in the neonatal Idua-W392X mouse brain, demonstrating potential for mitigation of brain disease in MPS-IH.

“These data suggest that C3.MPS LNPs facilitate partial biochemical correction of disease in the neonatal Idua-W392X mouse brain, demonstrating potential for mitigation of brain disease in MPS-IH.”

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

The macaque experiments were conducted at the SingHealth Experimental Medicine Centre, accredited by the Association for Assessment and Accreditation of Laboratory Animal Care International.

“The macaque experiments were conducted at the SingHealth Experimental Medicine Centre, accredited by the Association for Assessment and Accreditation of Laboratory Animal Care International.”

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

In addition to the identity of the ionizable lipid, a range of variables during LNP formulation are known to influence in vivo delivery of gene editing platforms.

“In addition to the identity of the ionizable lipid, a range of variables during LNP formulation are known to influence in vivo delivery of gene editing platforms.”

May combine two claims.

The improvement in IDUA and GAG levels is likely due to secretion of IDUA enzyme into the CNS by a small number of corrected periventricular cells, which is supported by studies in neonatal reporter mice demonstrating the predominance of editing confined to periventricular cells after intraventricular injection of LNPs.

“The improvement in IDUA and GAG levels is likely due to secretion of IDUA enzyme into the CNS by a small number of corrected periventricular cells, which is supported by our studies in neonatal reporter mice demonstrating the predominance of editing confined to periventricular cells after intraventricular injection of LNPs.”

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

All human specimens were collected following Institutional Review Board (IRB) approval at the Children's Hospital of Philadelphia (IRB 21-018470).

“All human specimens were collected following Institutional Review Board (IRB) approval at the Children’s Hospital of Philadelphia (IRB 21-018470).”

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

All mouse animal protocols were approved by the Institutional Animal Care and Use Committee (IACUC) at Children's Hospital of Philadelphia (IACUC 21-001417) and followed guidelines in the NIH's Guide for the Care and Use of Laboratory Animals.

“All mouse animal protocols were approved by the Institutional Animal Care and Use Committee (IACUC) at Children’s Hospital of Philadelphia (IACUC 21-001417) and followed guidelines in the NIH’s Guide for the Care and Use of Laboratory Animals.”

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

While the lead LNP engineered in this study holds promise for CNS diseases in which a paracrine effect of restoring normal protein function following periventricular cell targeting is adequate, further LNP optimization is required to apply this delivery modality to a broader range of diseases involving non-periventricular structures or specific neural cell types.

“Thus, while the lead LNP engineered in this study holds promise for CNS diseases in which a paracrine effect of restoring normal protein function following periventricular cell targeting is adequate, further LNP optimization is required to apply this delivery modality to a broader range of diseases involving non-periventricular structures or specific neural cell types.”

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

Macaque procedures were performed in accordance with the IACUC at the National University of Singapore and Singapore Health Services (IACUC 2009-SHS-512).

“Macaque procedures were performed in accordance with the IACUC at the National University of Singapore and Singapore Health Services (IACUC 2009-SHS-512).”

Classified as probably not a substantial claim.

All quantifications represent the mean and standard error of the mean (SEM) acquired from at least three biological replicates (cells or animals) per treatment group.

“All quantifications represent the mean and standard error of the mean (SEM) acquired from at least three biological replicates (cells or animals) per treatment group.”

May combine two claims.

The engineered LNP platform demonstrated efficacy and safety in a mouse model of a human disease, in the fetal NHP model, and in patient-derived brain tissues.

“Here, we engineer an LNP platform to begin to address this need, demonstrating its efficacy and safety in a mouse model of a human disease, in the fetal NHP model, and in patient-derived brain tissues.”

• AssumptionRests on an assumption: The Idua-W392X mouse model recapitulates human MPS-IH.