IP Library › Granted Patent US 12,428,462
Granted Patent B2
US 12,428,462 · App. 18/306,911 · Granted Sep 30, 2025

Recombinant CDHR3 protein fragments inhibit rhinovirus C binding and replication

Inventors: Ann Carol Palmenberg (Madison, WI); Kelly Watters (Madison, WI)
Assignee: WISCONSIN ALUMNI RESEARCH FOUNDATION
C07K14/705G01N33/56983A61K38/00
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Quick Facts
Patent No.
US 12,428,462
App. No.
18/306,911
Granted
Sep 30, 2025
Kind
B2
Abstract

The present invention provides soluble truncated peptides of CDHR3, recombinant variants thereof and methods of making these peptides. The present invention also provide methods of inhibiting rhinovirus C infection and an in vitro assay for screening for anti-viral agents against rhinovirus C.

Claims (27)

1. A soluble truncated CDHR3 peptide comprising:

(a) domain 1 (SEQ ID NO: 2) of CDHR3 and linker 3 (SEQ ID NO: 17) positioned after domain 1 or

(b) domain 1 of CDHR3 with a mutation at position 76 relative to SEQ ID NO: 1 (SEQ ID NO: 31), wherein X is selected from the group of amino acids consisting of A, G, V, L, I, S, and T and linker 3 (SEQ ID NO: 17) positioned after domain 1.

2. The soluble truncated CDHR3 peptide of claim 1 , wherein the peptide consists of SEQ ID NO: 27 or consists of a peptide having at least 90% identity to SEQ ID NO: 27.

3. The soluble truncated CDHR3 peptide of claim 1 , wherein the peptide is covalently or non-covalently linked to a heterologous tag.

4. The soluble truncated CDHR3 peptide of claim 3 , wherein the tag is a FLAG tag or a HIS tag.

5. The soluble truncated CDHR3 peptide of claim 1 , wherein the soluble truncated CDHR3 peptide does not encompass full length CDHR3 (SEQ ID NO: 1).

6. The soluble truncated CDHR3 peptide of claim 1 , wherein the soluble truncated CDHR3 peptide does not include the sequences of domains 4, 5, and 6 of wild type CDHR3 (SEQ ID NO: 1).

7. The soluble truncated CDHR3 peptide of claim 1 , wherein the soluble truncated CDHR3 peptide encompasses 350 amino acids or less of the full length CDHR3 (SEQ ID NO: 1).

8. The soluble truncated CDHR3 peptide of claim 1 , further comprising:

(c) an additional sequence comprising

domain 3 of CDHR3 (SEQ ID NO: 4); and

(d) at least one additional linker, wherein the at least one linker is before domain 1or after domain 3, wherein the linkers are selected from:

(i) linker 1 (SEQ ID NO:15) before domain 1,

(ii) linker 4 (a peptide consisting of Pro-Ala-Thr-Xaa) positioned after domain 3, wherein Xaa is selected from the group consisting of A, G, V, L, I, S, and T, and

(v) a combination of any of (1)-(ii).

9. The soluble truncated CDHR3 peptide of claim 8 , wherein

linker 3 (SEQ ID NO: 17) is positioned between domain 1 and 3.

10. The soluble truncated CDHR3 peptide of claim 8 , wherein the peptide comprises domain 1 (SEQ ID NO: 2) and at least linker 4, wherein linker 4 is a peptide consisting of Pro-Ala-Thr-Xaa, wherein Xaa is selected from the group consisting of A, G, V, L, I, S, and T.

11. A vector comprising the nucleic acids encoding the soluble truncated CDHR3 peptide of claim 1 , wherein the nucleic acids comprise SEQ ID NO:14 (EC1+3 (Δ2)).

12. A cell able to express the soluble truncated CDHR3 peptide of claim 1 .

13. A therapeutic composition for reducing or preventing Rhinovirus C entry into cells, the composition comprising the soluble truncated peptide of claim 1 and Ca++in a pharmaceutically acceptable carrier.

14. A method for reducing an infection by human rhinovirus (HRV)-C of a host cell susceptible to infection by HRV-C, comprising:

contacting the virus with the soluble truncated peptide of claim 1 in an amount effective to reduce the infectivity of the virus.

15. An in vitro assay for testing an agent for anti-viral activity against rhinovirus C, the assay comprising the steps of:

(a) contacting the agent with the soluble truncated peptide of claim 1 and rhinovirus C; and

(b) assaying the ability of the agent to disrupt binding of the soluble truncated peptide of claim 1 to the virus.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 28, 2023
From: PALMENBERG, ANN; WATTERS, KELLY
To: WISCONSIN ALUMNI RESEARCH FOUNDATION
Reel/Frame 063479/0224 →
Continuity (4)
Continuation 17058934
Provisional Application 62768191 · Nov 16, 2018
Provisional Application 62678507 · May 31, 2018
Related Publication 20240109948A1 · Apr 4, 2024
References Cited (72)
US 9938507B2 · Gern et al. · 2018 [cited by applicant]
US 10280405B2 · Gern et al. · 2019 [cited by applicant]
US 20020042386A1 · Rosen · 2002 [cited by examiner]
US 20150064236A1 · Bancel · 2015 [cited by examiner]
US 20170082609A1 · Gern · 2017 [cited by examiner]
WO 2002044340A2 · 2002 [cited by applicant]
Ota et al., 2004, Complete sequencing and characterization of 21,243 full-length humans cDNAs, Nature Genetics, 36(1): 40-45. [cited by examiner]
Hillier et al., 2003, The DNA sequence of human chromosome 7, Nature, 424: 157-164. [cited by examiner]
Perez, T. D., et al. “Cadherin adhesion: mechanisms and molecular interactions.” Cell adhesion. Springer, Berlin, Heidelberg, 2004. 3-21. [cited by applicant]
Plummer THJ, et al (1984) Demonstration of peptide:N-glycosidase F activity in endo-beta-N-acetylglucosaminidase F preparations. J Biol Chem 259: 10700-10704. [cited by applicant]
Pokutta, S., et al., Conformational changes of the recombiant extracellular domain of E-cadherin upon calcium binding. Eur J Biochem, 1994. 223: p. 1019-1026. [cited by applicant]
Quist AP, et al (2000) Physiological role of gap-junctional hemichannels. Extracellular calcium-dependent isosmotic volume regulation. J Cel Biol 148: 1063-1074. [cited by applicant]
Shapiro L, et al. “Structure and biochemistry of cadherins and catenins.” Cold Spring Harbor perspectives in biology 1.3 (2009): a003053. [cited by applicant]
Shapiro L., et al., Structural basis of cell-cell adhesion by cadherins. Nature, 1995. 374: p. 327-337. [cited by applicant]
Sheehy AM, et al (2002) Isolation of a human gene that inhibits HIV-1 infection as is supressed by the viral Vif protein. Nature 418: 646-650. [cited by applicant]
Sherer, N.M., et al., Visualization of retroviral replication in living cells reveals budding into multivesicular bodies. Traffic., 2003. 4: p. 785-801. [cited by applicant]
Suzuki, Y., Sialobiology of influenza molecular mechanisms of host range variation of influenza viruses. Biological and Pharmaceutical Bulletin, 2004. 28: p. 399-408. [cited by applicant]
Troyanovsky, R.B., et al., Adhesisve and lateral E-cadherin dimers are mediated by the same interface. Mol and Cell Biol., 2003. 23: p. 7965-7972. [cited by applicant]
Turunen R, et al. (2016) Rhinovirus species and clinical characteristics in the first wheezing episode in children. J Med Virol 88: 2059-2068. [cited by applicant]
Watters et al., 2018, CDH R3 extracellular domains EC1-3 mediate rhinovirus C interaction with cells and as recombinant derivatives, are inhibitory to virus infection, PLoS Pathogens, 14(12): e1007 477 (21 pages). [cited by applicant]
Wheelock MJ, et al (2003) Cadherin-mediated cellular signaling. Curr Op Cell Biol 15: 509-514. [cited by applicant]
Yanai I, et al. (2005) Genome-wide midrange transcription profiles reveal expression level relationships in human tissue specification. Bioinformatics 21: 650-659. [cited by applicant]
Anonymous. CR201_G0018053-CDHR3 isoform 7— [cited by applicant]
Aricescu AR, et al. (2006) A time- and cost-efficient system for high-level protein production in mammalian cells. Acta Crystallogr D Biol Crystallogr 62: 1243-1250. [cited by applicant]
Ashraf S, et al (2013) Biological characteristics and propagation of human rhinovirus-C in differentiated sinus epithelial cells. Virology 436: 143-149. [cited by applicant]
Bizzintino J, et al. (2011) Association between human rhinovirus C and severity of acute asthma in children. Eur Respir J 37: 1037-1042. [cited by applicant]
Bochkov YA, et al. (2011) Molecular modeling, organ culture and reverse genetics for a newly identified human rhinovirus C. Nature Medicine 17: 627-632. [cited by applicant]
Bochkov YA, et al. (2015) Cadherin-related family member 3, a childhood asthma susceptibility gene product, mediates rhinovirus C binding and replication. Proc Natl Acad Sci USA 112: 5485-5490. [cited by applicant]
Bochkov YA, et al., Mutations in VP1 and 3A proteins improve binding and replication of rhinovirus C15 in HeLa-E8 cells. Virology, 2016. 499: p. 350-360. [cited by applicant]
Boggon, T.J., et al., C-cadherin ectodomain structure and implications for cell adhesion mechanisms. Science, 2002. 296: p. 1308-1313. [cited by applicant]
Bonnelykke K, et al. (2014) A genome-wide association study identifies CDHR3 as a susceptibility locus for early childhood asthma with severe exacerbations. Nature Genetics 46: 51-55. [cited by applicant]
Bonnelykke, K., et al. “Cadherin-related family member 3 genetics and rhinovirus C respiratory illnesses.” American journal of respiratory and critical care medicine 197.5 (2018): 589-594. [cited by applicant]
Chen, X. et al, Advances in the biology and chemistry of sialic acids. ACS Chemical Biology, 2011. 5: p. 163-176. [cited by applicant]
Cox DW, et al. (2013) Human rhinovirus species C infection in young children with acute wheeze is associated with increased acute repiratory hospital admissions. Am J Respir Crit Care Med 188: 1358-1364. [cited by applicant]
Database Geneseq. “Human extracellular matrix protein from gene 18.” Dec. 18, 2003. XP002793520, retrieved from EBI accession No. GSP:ADC10825 Database accession No. ADC10825. [cited by applicant]
Database Geneseq. “Human novel polypeptide #69.” Aug. 30, 2002. XP002793521, retrieved from EBI accession No. GSP:ABG66734 Database accession No. ABG66734. [cited by applicant]
Dominguez SR, et al. (2008) Multiplex MassTag PCR for respiratory pathogens in pediatric nasopharyngeal washes by conventional diagnostic testing shows a high prevalence of viruses belonging to a newly recognized rhinov… [cited by applicant]
Gallois-Montbrun S, et al. (2007) Antiviral protein APO-BEC3G localizes to ribonucleoprotein complexes found in P bodies and stress granules. J Virol 81: 2165-2178. [cited by applicant]
Gern JE (2010) The ABCs of rhinoviruses, wheezing, and asthma. J Virol 84: 7418-7426. [cited by applicant]
Greve JM, et al. (1989) The major human rhinovirus receptor is ICAM-1. Cell 56: 839-847. [cited by applicant]
Greve JM, et al. Mechanisms of receptor-mediated rhinovirus neutralization defined by two soluble forms of ICAM-1. J Virol, 1991. 65: p. 6015-6023. [cited by applicant]
Griggs TF, et al. (2015) Production, purification, and capsid stability of rhinovirus C types. J Virol Methods 217: 18-23. [cited by applicant]
Griggs TF, et al. (2017) Rhinovirus C targets ciliated airway epithelial cells. Respiratory Research 18: 84. [cited by applicant]
Guo, H., et al., Regulation of homotypic cell-cell adhesion by branched N-glycosylation of N-cadherin extracellular EC2 and EC3 domains. J Biol Chem, 2009. 284: p. 34986-34997. [cited by applicant]
Halbleib JM, et al. (2006) Cadherins in development: cell adhesion, sorting, and tissue morphogenesis. Genes and Dev 20: 3199-3214. [cited by applicant]
Hao W, et al. (2012) Infection and propagation of human rhino-virus C in human airway epilethial cells. J Virol 86: 24-32. [cited by applicant]
Harrison, O.J., et al., The extracellular architecture of adherens junctions revealed by crystal structures of type I cadherins. Struct., 2011. 19: p. 244-256. [cited by applicant]
Haussinger, D., et al., Calcium-dependent homoassociation of Eclient cadherin by NMR Spectroscopy: changes in mobility, conformation and mapping of contact regions. J Mol Biol, 2002. 324: p. 823-839. [cited by applicant]
Hofer F, et al. (1994) Members of the low density lipoprotein receptor family mediate cell entry of a minor-group common cold virus. Proc Natl Acad Sci U S A 91: 1839-1842. [cited by applicant]
International Searching Authority. International Search Report and Written Opinion for application PCT/US2019/034580. Mailed on Sep. 6, 2019. 16 pages. [cited by applicant]
Jartti T, et al. (2009) Bronchiolitis: age and previous wheezing episodes are linked to viral etiology and atopic characteristics. Pediatr Infect Dis J 28: 311-317. [cited by applicant]
Juven T, et al. (2000) Etiology of community-acquired pneumonia in 254 hospitalized children. Pediatr Infect Dis J 19: 293-298. [cited by applicant]
Kartenbeck J, et al (1991) Endocytosis of junctional cadherins in bovine kidney epithelial (MDBK) cells cultured in low Ca2+ ion medium. J Cel Biol 113: 881-892. [cited by applicant]
Kim D, et al. (2004) Protein structure prediction and analysis using the Robetta server. Nucleic Acid Res 32: 526-531. [cited by applicant]
Koch, A.W., et al., Calcium binding and homoassociation of E-cadherin domains. Biochemistry, 1997. 36: p. 7697-7705. [cited by applicant]
Langer, M.D., et al., N-glycosylation alters cadherin-mediated intercellular binding kinetics. J Cell Sci, 2012. 125: p. 2478-2485. [cited by applicant]
Last-Barney, K., et al., Detection of major group rhinoviruses by soluble intercellular adhesion molecule-1 (SICAM-1). Jornal of Virological Methods, 1991. 35: p. 255-264. [cited by applicant]
Lee W-M, et al. (2012) Human rhinovirus species and season of infection determine illness severity. Am J Respir Crit Care Med 186: 886-891. [cited by applicant]
Liu, Y., et al. (2015) Sialic acid-dependent cell entry of human enterovirus D68. Nature Communications. [cited by applicant]
Liu, Y., et al., Atomic structure of a rhinovirus C, a virus species linked to severe childhood asthma. Proc. Nat. Acad. Sci. USA, 2016. 113: p. 8997-9002. [cited by applicant]
Liu, Y., et al., Structure and inhibition of EV-D68, a virus that causes respiratory illness in children. Science, 2015. 347: p. 71-74. [cited by applicant]
Liwosz A, et al. (2006) N-glycosylation affects the molecular organization and stability of E-cadherin junctions. J Biol Chem 281: 23138-23149. [cited by applicant]
Marlovits, T.C., et al., Very-low-density lipoprotein receptor fragment shed from HeLa cells inhibits human rhinovirus infection. J Virol, 1998. 72: p. 10246-10250. [cited by applicant]
Mattey DL, et al (1986) Splitting and internalization of the desmosomes of cultured kidney epithelial cells by reduction in calcium concentration. J Cell Sci 85: 113-124. [cited by applicant]
McErlean P, et al. (2007) Characterisation of a newly identified human rhinovirus, HRV-QPM, discovered in infants with bronchiolitis. J Clin Virol 39: 67-75. [cited by applicant]
Nagar, B., et al., Structural basis of calcium-induced E-cadherin rigidification and dimerization. Nature, 1996. 380: p. 360-364. [cited by applicant]
Nakagome, K., et al., Effects of rhinovirus species on viral replication and cytokine production. J Allergy Clin Immunol, 2014. 134: p. 332-341. [cited by applicant]
Nejsum LN, et al. (2007) A molecular mechanism directly linking E-cadherin adhesion to initiation of epithelial cell surface polarity. J Cel Biol 178: 323-335. [cited by applicant]
Niessen, C.M., et al., Tissue organization by cadherin adhesion molecules: Dynamic molecular and cellular mechanisms of morphogenetic regulation. Physiol Rev., 2011. 91: p. 691-731. [cited by applicant]
O'Doherty U, et al.(2000) Human immunodeficiency virus type 1 spinoculation enhances infection through virus binding. J Virol 74: 10074-10080. [cited by applicant]
O'Neill MB, et al.(2017) Evolutionary genetics of a disease suceptibility locus in CDHR3. bioRxiv http://dx.doi.org/10.1101/186031. [cited by applicant]
Palmenberg, A.C., Rhinovirus C, asthma, and cell surface expression of virus receptor, CDHR3. J Virol, 2017. 91. [cited by applicant]