US 2008005.7509A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2008/0057509 A1 Lupski et al. (43) Pub. Date: Mar. 6, 2008 (54) DEFECTS IN PERIAXIN ASSOCIATED WITH (60) Provisional application No. 60/255.217, filed on Dec. MYELINOPATHIES 13, 2000. (76) Inventors: James R. Lupski, Houston, TX (US); Publication Classification Cornelius F. Boerkoel III, Houston, TX (US); Hiroshi Takashima, Houston, (51) Int. Cl. TX (US) CI2O I/68 (2006.01) (52) U.S. Cl. .................................................................. 435/6 Correspondence Address: FULBRIGHT & JAWORSKI, LLP 1301 MCKNNEY SUTE 51OO (57) ABSTRACT HOUSTON, TX 77010-3095 (US) (21) Appl. No.: 11/838,500 The present invention relates to defects in periaxin (PRX) associated with myelinopathies, including Charcot-Marie (22) Filed: Aug. 14, 2007 Tooth syndrome and/or Deerine-Sottas syndrome. Unre lated individuals having a myelinopathy from Dejerine Related U.S. Application Data Sottas syndrome have recessive PRX mutations. The PRX locus maps to a region associated with a severe autosomal (63) Continuation of application No. 10/021,955, filed on recessive demyelinating neuropathy and is also syntenic to Dec. 13, 2001, now Pat. No. 7,273,698. the Prx location on murine chromosome 7. PN-44. v Fifi is Gf * is f. f i? it it it it i? it is 86 if f : Gift it if is f. 6 it is a a is 8 is a B g g it a fit i AN AIK hetero 45A homoliSA homoASA CSX 7 Six Taar a via a unania t fi : Yi 3 y Clas?.(CENELY I II. Ex. AA.y X. | eterodec hetero.247del PN-6. PN-7s2 PN-76.3 PN-76.4 N-76.5 PN-766 * a a a 3 4:3, 8: 83 8:38.8 : is a 3 a 3 3 A . AAAIA, A.AA. AA AY; X& AA- All Nifi A: AAAEY s ly.a nav is in MISL honoave hetero247 dec Wildtype hetero.247deC RB2sses Patent Application Publication Mar. 6, 2008 Sheet 1 of 9 US 2008/005.7509 A1 Patent Application Publication Mar. 6, 2008 Sheet 3 of 9 US 2008/005.7509 A1 HOU 297 2857C>T is 5 GCI SS GN A S Ct a C a Patient 851 3 : HOU 579 46 HOU 418 36 2787AC C C C Patient 136 Patent Application Publication Mar. 6, 2008 Sheet 4 of 9 Patent Application Publication Mar. 6, 2008 Sheet 5 of 9 US 2008/005.7509 A1 Patent Application Publication Mar. 6, 2008 Sheet 6 of 9 US 2008/005.7509 A1 Patent Application Publication Mar. 6, 2008 Sheet 7 of 9 US 2008/005.7509 A1 Patent Application Publication Mar. 6, 2008 Sheet 8 of 9 US 2008/005.7509 A1 Patent Application Publication Mar. 6, 2008 Sheet 9 of 9 US 2008/005.7509 A1 US 2008/005.7509 A1 Mar. 6, 2008 DEFECTS IN PERAXN ASSOCATED WITH tating integration of extracellular signaling through the MYELINOPATHES cytoskeleton which is essential for changes in Schwann cell shape and regulation of gene expression during axonal 0001. The present invention claims priority to U.S. Non ensheathment (Fernandez-Valle et al., 1997; Tapon and Hall, provisional patent application Ser. No. 10/021,955, filed 1997). Such a signaling function is supported by the obser Dec. 13, 2001, and U.S. Provisional Patent Application Vation that L-periaxin contains a PDZ motif, a domain 60/255,217, filed Dec. 13, 2000, both of which are incor implicated in the assembly of signaling complexes at sites of porated by reference herein in their entirety. cell-cell contact, and a nuclear localization signal (Dytrych 0002 The present invention was developed with funds et al., 1998: Sherman and Brophy, 2000). Confirming the from the United States Government. Therefore, the United necessity of periaxin for maintenance of the myelin sheath, States Government may have certain rights in the invention. Gillespie et al. recently demonstrated that Prx mice ensheathe and myelinate peripheral axons apparently nor FIELD OF THE INVENTION mally but Subsequently develop a severe demyelinating 0003. The present invention is directed to the fields of neuropathy associated with allodynia (pain from non-nox molecular biology, molecular genetics, and neurology. More ious stimuli) and hyperalgesia (hypersensitivity to pain) specifically, the present invention is directed to defects in (Gillespie et al., 2000). periaxin related to neuropathies. More specifically, the neu 0006. However, it was heretofore unknown in the art ropathies include recessive Deerine-Sottas and Charcot whether a relationship between the human PRX gene defects Marie-Tooth disease. and neuropathies such as recessive DSN existed. BACKGROUND OF THE INVENTION SUMMARY OF THE INVENTION 0007. In an embodiment of the present invention there is 0004 Dejerine-Sottas neuropathy (DSN) and Charcot a method of diagnosing myelinopathy in an individual Marie-Tooth disease type 1 (CMT1) represent genetically comprising the steps of obtaining a sample containing heterogeneous inherited peripheral myelinopathies. These nucleic acid from the individual; assaying the sample for an conditions constitute part of a spectrum of neuropathy alteration in a periaxin polynucleotide, wherein the alter phenotypes ranging in severity from congenital hypomyeli ation is associated with the myelinopathy. In a specific nating neuropathy (CHN) to adult onset hereditary neuropa embodiment, the myelinopathy is selected from the group thy with liability to pressure palsies (HNPP) (Lupski and consisting of Charcot-Marie-Tooth (CMT) syndrome, Garcia, 2001). At least fifteen genetic loci and six genes have hereditary neuropathy with liability to pressure palsies been associated with these disorders; identified genetic causes include altered dosage of peripheral myelin protein (HNPP), Dejerine-Sottas syndrome (DSS), congenital hypo 22 (PMP22) or mutations in one of the following genes: myelinating neuropathy (CHN), and Roussy-Levy Syn PMP22, the gap junction protein B1 gene (GJB1), the myelin drome (RLS). In another specific embodiment, the assaying protein Zero gene (MPZ), the early growth response gene 2 step further comprises a polymerase chain reaction. In a (EGR2), the myotubularin related protein 2 gene (MTMR2), further specific embodiment, the primers for said poly or the N-myc downstream regulated gene 1 (NDRG1)) merase chain reaction are selected from the group consisting (Lupski and Garcia, 2001). These genes encode proteins of of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID diverse functions: compact myelin structural proteins (MPZ. NO:6, SEQID NO:7, SEQID NO:8, SEQID NO:9, SEQID PMP22), a non-compact myelin gap junction protein NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, (GJB1), signal transduction proteins (NDRG1, MTMR2), SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID and a transcription factor for late myelin genes (EGR2). NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, Both dominant (PMP22, GJB1, MPZ, EGR2) and recessive SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID (MTMR2, NDRG1, PMP22, EGR2) mutant alleles have NO:24, SEQ ID NO:25, SEQ ID NO:26. In a specific been described. Historically considered an autosomal reces embodiment, the alteration is 3775G>A, 1216G>A, 4075 sive disorder (Dejerine and Sottas, 1893), DSN has been 4077d, 1483G>C, 3394A-G, 3248C>G, 2763 A-G, associated predominately with de novo dominant mutations 2645C>T, 306C>T, 1491C>G, or 2655T>C. in PMP22 (Roa et al., 1993), MPZ (Hayasaka et al., 1993), 0008. In another embodiment of the present invention or EGR2 (Timmerman et al., 1999), although rare recessive there is a method of diagnosing myelinopathy in an indi mutations in PMP22 have also been reported (Lupski, 2000; vidual comprising the steps of obtaining a sample containing Parman et al., 1999). protein from the individual; assaying the sample for an 0005. In murine embryonic Schwann cells, L-periaxin is alteration in a periaxin polypeptide, wherein the alteration is initially concentrated in the nuclei but redistributes to the associated with the myelinopathy. In a specific embodiment, plasma membrane, predominantly adaxonal, with initiation the alteration is E1259K, A406T, E1359delA, E495Q, of myelination and then to the abaxonal, Schmidt-Lanter R1132G, P1083R, 1921M, A882V, T102T, P497P, P885P, man incisures, and paranodal membranes with maturation of R953X, R368X, S929fsX957, R196X, V763fsX774, the myelin sheath (Scherer et al., 1995; Sherman and Bro C715X, or R82fsX96. In another specific embodiment, the phy, 2000). In addition, L-periaxin expression recapitulates myelinopathy is selected from the group consisting of Char this pattern following crush injury (Scherer et al., 1995). cot-Marie-Tooth (CMT) syndrome, hereditary neuropathy This shift in periaxin localization after the spiralization with liability to pressure palsies (HNPP), Dejerine-Sottas phase of myelination suggests that periaxin participates in syndrome (DSS), congenital hypomyelinating neuropathy membrane-protein interactions that are required to stabilize (CHN), and Roussy-Levy Syndrome (RLS). the mature myelin sheath. As a cytoskeleton-associated 0009. In another embodiment of the present invention protein, L-periaxin may mediate Such stabilization by facili there is as a composition of matter a defect of a periaxin US 2008/005.7509 A1 Mar. 6, 2008 polynucleotide of 3775GdA, 1216G>A, 4075-4077d, comprising the step of administering to the organism a 1483G>C, 3394A-G, 3248C>G, 2763A-G, 2645C>T, therapeutically effective amount of a periaxin nucleic acid 306C>T, 1491C>G, and 2655T>C. sequence, wherein the nucleic acid sequence is administered 0010. In an additional embodiment of the present inven by a vector. In a specific embodiment, the vector is selected tion there as a composition of matter a periaxin polypeptide from the group consisting of a plasmid, a viral vector, a lipid, defect of E1259K, A406T, E1359delA, E495Q, R1132G, a liposome, a polypeptide, or a combination thereof.