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Mucopolysaccharidosis I (MPS I), a lysosomal storage disease caused by dysfunction of α-L-iduronidase (IDUA), is characterized by the deposition of dermatan sulfate (DS) and heparan sulfate (HS) throughout the body, which causes several somatic and central nervous symptoms. Although enzyme-replacement therapy (ERT) is currently available to treat MPS I, it does not alleviate central nervous disorders, as it cannot penetrate the blood-brain barrier. Here we evaluate the brain delivery, efficacy, and safety of JR-171, a fusion protein comprising humanized anti-human transferrin receptor antibody Fab and IDUA, using monkeys and MPS I mice. Intravenously administered JR-171 was distributed in major organs, including the brain, and reduced DS and HS concentrations in the central nervous system and peripheral tissues. JR-171 exerted similar effects on peripheral disorders similar to conventional ERT and further reversed brain pathology in MPS I mice. We found that JR-171 improved spatial learning ability, which was seen to deteriorate in the vehicle-treated mice. Further, no safety concerns were noted in repeat-dose toxicity studies in monkeys. This study provides nonclinical evidence that JR-171 might potentially prevent and even improve disease conditions in patients with neuronopathic MPS I without serious safety concerns., Competing Interests: S.K., Y.K., E.Y., A.Y., H.M., A. Imakiire, N.T., S.T., A.M., J.I., A. Inoue, R.Y., K.M., T.H., K.T., and H.S. are employees and/or stockholders of JCR Pharmaceuticals Co., Ltd., (© 2023 The Author(s).)

Mucopolysaccharidosis II (MPS II), a lysosomal storage disease caused by mutations in iduronate-2-sulfatase (IDS), is characterized by a wide variety of somatic and neurologic symptoms. The currently approved intravenous enzyme replacement therapy with recombinant IDS (idursulfase) is ineffective for CNS manifestations due to its inability to cross the blood-brain barrier (BBB). Here, we demonstrate that the clearance of heparan sulfate (HS) deposited in the brain by a BBB-penetrable antibody-enzyme fusion protein prevents neurodegeneration and neurocognitive dysfunctions in MPS II mice. The fusion protein pabinafusp alfa was chronically administered intravenously to MPS II mice. The drug reduced HS and attenuated histopathological changes in the brain, as well as in peripheral tissues. The loss of spatial learning abilities was completely suppressed by pabinafusp alfa, but not by idursulfase, indicating an association between HS deposition in the brain, neurodegeneration, and CNS manifestations in these mice. Furthermore, HS concentrations in the brain and reduction thereof by pabinafusp alpha correlated with those in the cerebrospinal fluid (CSF). Thus, repeated intravenous administration of pabinafusp alfa to MPS II mice decreased HS deposition in the brain, leading to prevention of neurodegeneration and maintenance of neurocognitive function, which may be predicted from HS concentrations in CSF., (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)

Mucopolysaccharidosis II (MPS II) is an X-linked recessive lysosomal storage disease caused by mutations in the iduronate-2-sulfatase (IDS) gene. Since IDS catalyzes the degradation of glycosaminoglycans (GAGs), deficiency in this enzyme leads to accumulation of GAGs in most cells in all tissues and organs, resulting in severe somatic and neurological disorders. Although enzyme replacement therapy with human IDS (hIDS) has been used for the treatment of MPS II, this therapy is not effective for defects in the CNS mainly because the enzyme cannot cross the blood-brain barrier (BBB). Here, we developed a BBB-penetrating fusion protein, JR-141, which consists of an anti-human transferrin receptor (hTfR) antibody and intact hIDS. The TfR-mediated incorporation of JR-141 was confirmed by using human fibroblasts in vitro. When administrated intravenously to hTfR knockin mice or monkeys, JR-141, but not naked hIDS, was detected in the brain. In addition, the intravenous administration of JR-141 reduced the accumulation of GAGs both in the peripheral tissues and in the brain of hTfR knockin mice lacking Ids, an animal model of MPS II. These data provide a proof of concept for the translation of JR-141 to clinical study for the treatment of patients with MPS II with CNS disorders., (Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.)

Multivesicular bodies (MVBs) are late endosomal compartments containing luminal vesicles (MVB vesicles) that are formed by inward budding of the endosomal membrane. In budding yeast, MVBs are an important cellular mechanism for the transport of membrane proteins to the vacuolar lumen. This process requires a class E subset of vacuolar protein sorting (VPS) genes. VPS44 (allelic to NHX1) encodes an endosome-localized Na(+)/H(+) exchanger. The function of the VPS44 exchanger in the context of vacuolar protein transport is largely unknown. Using a cell-free MVB formation assay system, we demonstrated that Nhx1p is required for the efficient formation of MVB vesicles in the late endosome. The recruitment of Vps27p, a class E Vps protein, to the endosomal membrane was dependent on Nhx1p activity and was enhanced by an acidic pH at the endosomal surface. Taken together, we propose that Nhx1p contributes to MVB formation by the recruitment of Vps27p to the endosomal membrane, possibly through Nhx1p antiporter activity.

Since rotavirus vaccines aim to protect children from severe diarrhea, knowledge of the prevailing G serotypes among rotaviruses from hospitalized children is essential. Thus, we determined the G serotypes of rotaviruses collected from children with acute diarrhea in a local referral hospital in Akita, Japan, over the 10-year period between January 1987 and December 1996. Based on the assumption that rotaviruses with an identical electropherotype possess the same G serotype, the G serotypes of 488 rotavirus-positive specimens that were classified into 63 electropherotypes were determined by enzyme-linked immunosorbent assay with a supplementary use of G typing by reverse transcription-PCR. The relative frequencies over the 10-year period were 77.0 (G1), 14.5 (G2), 2.7 (G3) and 5.3% (G4), leaving the possibility that only 0.4% had G serotypes uncommon to human rotaviruses. Of 24,050 rotaviruses extracted by reviewing 63 serotyping studies in literature, the relative frequencies of the four major G serotypes were 50.6 (G1), 9.3 (G2), 7.2 (G3) and 11.6% (G4). As to uncommon G serotypes, only 0.9% were described as serotypes other than G1-4, and our estimate for potential uncommon serotypes were at most 8.1%. Thus, both this long-term study focusing on the rotaviruses only from severe cases in a single hospital in Japan and the global review of G serotypes published to date indicate that the primary target of any rotavirus vaccines should be rotaviruses possessing serotypes G1-4.

While an increasing number of studies suggest that there is a high prevalence of rotaviruses with P8[11], a typical P type of bovine rotavirus (BRV), among human neonates or infants in India, no data are available on the distribution of G and P types of Indian BRVs. Thus, fecal specimens were collected from cow and buffalo calves under 1 month of age on organized dairy farms in India during the period between 1994 and 1997, and 36 rotavirus-positive specimens were used to determine the relative frequencies of the G and P types of Indian BRVs. As to the G type, G10 was predominant (83%), followed by G6 (6%). The majority (94%) of BRVs had P8[11], and only one isolate possessed P6[1]. The most common combination of G and P types was G10P8[11] (81%), followed by G6P6[1] (3%) and G6P8[11] (3%). The high prevalence of BRVs possessing P8[11] VP4s strongly supports the hypothesis that BRVs may cross the host species barrier and circulate among neonates in India.

Super-short rotavirus strains that have a rearranged gene segment 11 are rarely found in humans, and only five isolates, all from Southeast Asia, have been described in the literature. We report the first isolation in Japan from an infant with severe diarrhea of a rotavirus possessing a super-short RNA pattern. This strain, designated AU19, had a G1 VP7 and is also the first isolate in Japan that possesses a P2[6] VP4. Furthermore, the P2[6] VP4 carried by AU19 was divergent in the hypervariable region of the amino acid sequence from the P2A[6] VP4s carried by asymptomatic neonatal strains or from the P2B[6] VP4 carried by porcine rotavirus strain Gottfried. Thus, AU19 is likely to represent a new VP4 subtype, which we propose to call P2C. Given the recent emergence of the P2[6] VP4s in India, Brazil, and the United States and the role of VP4 in protective immunity, further scrutiny is justified to see whether the emergence of the previously underrepresented P2[6] VP4 serotype is related to this new P2 subtype.