Isolation and characterization of Hermansky Pudlak Syndrome...

Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues

Reexamination Certificate

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C514S002600, C435S317100

Reexamination Certificate

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06573364

ABSTRACT:

FIELD OF THE INVENTION
The present invention disclosed herein relates to complexes of the Hermansky-Pudlak Syndrome (HPS) protein with other proteins including, but not limited to, complexes of HPS with: 14-3-3 eta, Hrs, BMK1 alpha kinase, CDK2, Nuclear factor NF90, Atrophin-1, DGS-1, HPIP1 and human HN1 homolog protein. The present invention further relates to antibodies specific for HPS complexes, and their use in, inter alia, screening, diagnosis, prognosis and therapy. The present invention further relates to the HPIP1 and human HN1 homolog protein nucleic acid, protein and derivatives, fragments and analogs thereof.
BACKGROUND OF THE INVENTION
(1) Hermansky-Pudlak Syndrome (HPS) and Chediak Higashi Syndrome (CHS)
Hermansky-Pudlak syndrome (HPS) is a genetic disorder characterized by defective lysosome-related organelles. Although the frequency of HPS is quite low in the general population, the frequency is markedly higher in certain genetically-isolated population groups. For example, HPS occurs in northwest Puerto Rico with a prevalence of 1 in 1800. In humans, HPS is characterized by the symptomatic triad of oculocutaneous albinsim, platelet dysfunction (i.e., mild to moderate bleeding diathesis), and ceroid deposition. Tissue accumulation of ceroid pigment (ceroid desposition) is considered to cause several serious complications, including progressive pulmonary fibrosis leading to death in the fourth or fifth decades. The primary defect in HPS involves affects on the contents and/or the secretion of several subcellular organelles, including lysosomes, melanosomes, and platelet-dense granules. See e.g., Swank, et al., 1998
. Pigment Cell Res
. 11:60-80; Erickson, 1997
. Proc. Natl. Acad. Sci. USA
94:8924-8925. The synthesis of pigmented melanosomes is compromised in HPS patients, wherein functional melanocytes are quantitatively reduced in number and/or are qualitatively abnormal, thus resulting in albinism of both dermal and keratinized tissues (e.g., the skin and hair). See e.g., Gardner, et al., 1997
. Proc. Natl. Acad. Sci. USA
94:9238-9243. The prolonged bleeding time associated with HPS is due to the lack of the storage organelles, platelet-dense granules, which are required for ADP-release and platelet aggregation. See e.g., Holmsen, et al., 1979
. Ann. Rev. Med
. 30:119-134. The pulmonary fibrosis and granulomatous colitis demonstrated in some cases of HPS is due to the accumulation of ceroid lipofuscin in lysosomes of reticuloendothelial cells, bone marrow and lung macrophages, gastrointestinal mucosal cell, and other cell types. See e.g., Harmon, et al., 1994
. J. Lab. Clin. Med
. 123:617-627.
Similar physiological abnormalities have been shown to occur in the pale ear (ep) mouse. See Lane & Green, 1967
. J. Heredity
58:17-20. Homozygous recessive ep mice have decreased skin and eye pigment at birth, as well as abnormal organelle function resulting in an increase of serum lysosomal enzymes. See Novak & Swank, 1979
. Genetics
92:189-204. Similar deleterious physiological symptomology was also described in a murine model of Chediak-Higashi Syndrome (CHS), within the beige mouse. CHS is, like HPS, an autosomal recessive genetic disorder. Symptoms in affected patients involve hypopigmentation, immunological deficiency, a bleeding tendency and a neurological disorder, probably due to protein sorting defects, especially in secretory lysosomes of granular cells. See Spritz, 1998, J. Clin. Immunol. 18:97-105. Chediak-Higashi syndrome proteins (LYST and LYST-2) have been identified in mouse and in man. LYST interacting proteins include 14-3-3 protein, HS1 (14-3-3 beta) protein, Hrs, BMK1 alpha kinase, KB07, Efs, OS9, casein kinase II beta SU, calmodulin, Troponin, Importin beta, Fte-1, estrogen-receptor related protein, Imogen 38, Atrophin-1, GBDR1, DGS-I, noHPSin (KIAA0607), OPA containing protein, M4 protein, LIP1 (Tcp-10 homolog), LIP2 (L17 homolog), LIP3 (Roaz protein homolog), LIP4 (hnRNP-e2 homolog), LIP5, LIP6 (Ns2-3 homolog), LIP7 (TCP10A homolog), LIP8 (KAP4L homolog), LIP9 (etr-1 homolog), and LIP10, as described in U.S. patent application Ser. No. 09/054,956.
The gene encoding a HPS-related protein has recently been cloned in both humans and mice. See Oh, et al., 1996
. Nat. Genet
. 14:300-306; Gardner, et al., 1997
. Proc. Natl. Acad. Sci. USA
94:9238-9243. Sequence analysis has predicted this polypeptide (hereinafter the “HPS protein”) to be a component of cytoplasmic organelles due to the presence of a putative transmembranal region. The mRNA encoded by the HPS gene in ep mice appears to be very widely expressed.
In summary, it is most likely that both human and murine HPS symptoms results from a defect in a protein which is required for the normal assembly, maturation, and/or structure of numerous, diverse subcellular organelles. Accordingly, HPS is likely to be centrally implicated in pathological processes, including but not limited to, oculocutaneous albinism, fibrotic lung disease, and various clotting, bleeding, and neurodegenerative disorders.
(2) HPS Protein-Interacting Proteins of the Present Invention
As previously discussed, HPS patients have been reported to suffer from several serious medical conditions, including oculocutaneous albinsim, a bleeding diathesis, and ceroid deposition, often accompanied by severe fibrotic lung disease and granulomatous colitis. However, despite the recognition of these HPS-associated syndromes, no curative therapeutic intervention currently exists for this disease. Currently, only symptomatic treatment can be offered. One potential area of difficulty involves a lack of understanding regarding the interaction of the HPS protein with other cellular proteins. The elucidation of these potential interactions may provide the means for the subsequent development of an diagnostic assay and/or a therapeutic modality for HPS and its associated diseases.
The following sections will discuss the various proteins which have been shown to interact with the HPS protein. These HPS protein-interacting-proteins hereinafter “HPS-IP” may be differentiated into proteins which are involved in signaling processes and protein trafficking (14-3-3 eta, Hrs, BMK1 alpha, CDK2, NF90), those proteins involved in neurodegenerative and developmental disorders (Atrophin I, DGS-1) and those previously-uncharacterized, novel proteins (HPIP1, HN1 homolog). Interestingly, five of these nine interacting proteins, or similar proteins, were found to interact as well with the LYST protein, as described by Nandabalan and Kingsmore in U.S. patent application Ser. No. 09/054,956 (Apr. 3, 1998): 14-3-3 protein, HS1 (14-3-3 beta) protein, Hrs, BMK1 alpha kinase, Atrophin-1, and DGS-I. CDK2, NF 90, HPIP1, and HN1 homolog were not found to interact with LYST. Table I provides an overview of all HPS interacting proteins and their interacting domains as disclosed in by the present invention.
It should be noted that the citation of a reference in this or in any other section of the specification should not be construed as an admission that such reference is prior art to the present invention disclosed herein.
(A) Proteins Involved Signaling Processes and Protein Trafficking
(i) 14-3-3 eta
The caHPSoxy-terminal region (starting at nucleotide 764) of the 14-3-3 protein eta isoform (GenBank Accession Number X80536; Ichimura-Ohshima, et al., 1992
. J. Neurosci. Res
. 31:600-605) was found to interact with HPS protein in this invention. The nucleotide and amino acid sequences of a Hermansky-Pudlak Syndrome Protein sequence provided in GenBank Accession Number U65676 are as follows:
   1 gggcgctgtg cgcgccgcga tccggtacgt gggcctccgg gctgtcccct ctgggggcga
(SEQ ID NO:5)

  61 tcctccctcc ggagcccccc ttcaaccctc ccggaagtga ggaccaggga tgctgtgctg

 121 ctctcccatg agccagtcac cgagtcggtc tgctgcagcc ctttctgaac ctctggccgt

 181 ctggatgctc cactgtgctt gccaagatga agtgcgtctt ggtggccact gagggcgcag

 241 aggtcctctt ctactggaca gatcaggagt ttgaagagag tctccggctg aagttcgggc

 301 agtcagagaa tgaggaagaa gagctccctg ccctggagga ccagctcagc accct

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