Methods of diagnosing and treating cd55 deficiency, hyperactivation of complement, angiopathic thrombosis and protein losing enteropathy (chaple), a newly identified orphan disease

Abstract

Disclosed herein are methods of diagnosing, and treating and/or preventing CD55-deficiency, hyperactivation of complement, angiopathic thrombosis and protein-losing enteropathy (CHAPLE). The method of diagnosing includes: providing a sample from a patient; performing an assay detecting at least one of at least one mutation in a DNA sequence of a CD55 gene, at least one mutation in a RNA sequence of a CD55 transcript, at least one mutation in a DNA sequence of a CD55 complementary-DNA (cDNA), CD55 protein, CD55 protein binding, complement deposition or combinations thereof; and diagnosing the patient with CHAPLE. The method of treating and/or preventing at least one symptom of CHAPLE includes: administering an effective amount of a composition comprising at least one complement inhibitor to a subject in need thereof, wherein the composition is effective in treating or preventing at least one symptom of CHAPLE. The disclosure further relates to compositions effective at treating and/or preventing CHAPLE.

1 . A method of diagnosing a patient with CD55 deficiency, hyperactivation of complement, angiopathic thrombosis and protein losing enteropathy (CHAPLE), the method comprising:
 providing a sample from a patient;   detecting at least one of:
 at least one mutation in a DNA sequence of a CD55 gene; 
 at least one mutation in a RNA sequence of a CD55 transcript; 
 at least one mutation in a DNA sequence of a CD55 complementary DNA (cDNA); 
 CD55 protein; 
 CD55 protein binding; 
 complement deposition; or 
 combinations thereof, wherein a mutation in the CD55 gene, transcript or cDNA, or absence or decrease in activity of CD55 protein is indicative of a patient at risk of developing or having CHAPLE; and 
   diagnosing the patient as having or not having CHAPLE.   
     
     
         2 . The method of  claim 1 , wherein the patient is diagnosed with CHAPLE when at least one of the following is detected: at least one mutation in a DNA sequence, an RNA sequence or a cDNA sequence of CD55 that results in a CD55 protein with substantially diminished functional activity, a CD55 protein with no functional activity, a lack of expression of CD55 protein (i.e., no CD55 protein expression), or a substantially diminished expression of CD55 protein; a CD55 protein with substantially diminished functional activity; a CD55 protein with no functional activity; a lack of expression of CD55 protein (i.e., no CD55 protein expression); a substantially diminished expression of CD55 protein; or combinations thereof. 
     
     
         3 . The method of  claim 1 , wherein the patient is diagnosed with CHAPLE when the patient has at least one CHAPLE related symptom and at least one of the following is detected: at least one mutation in a DNA sequence, an RNA sequence or a cDNA sequence of CD55 that results in a CD55 protein with substantially diminished functional activity, a CD55 protein with no functional activity, a lack of expression of CD55 protein (i.e., no CD55 protein expression), or a substantially diminished expression of CD55 protein; a CD55 protein with substantially diminished functional activity; a CD55 protein with no functional activity; a lack of expression of CD55 protein (i.e., no CD55 protein expression); a substantially diminished expression of CD55 protein; or combinations thereof. 
     
     
         4 . The method of  claim 3 , wherein the CHAPLE related symptom is selected from the group consisting of: inflammatory bowel disease, enteropathy, protein losing enteropathy, protein losing enteropathy associated with hypoalbuminemia, hypoalbuminemia, hypogammaglobulinemia, intestinal lymphangiectasia, lymphangiectasia, thrombotic events, thromboembolism, hyperactivation of complement, angiopathic thrombosis, hypoproteinemia, or combinations thereof. 
     
     
         5 . The method of  claim 1 , further comprising administering an effective amount of a composition comprising at least one complement inhibitor to the subject with CHAPLE, wherein the composition is effective in treating or preventing at least one symptom of CHAPLE, wherein the complement inhibitor is selected from the group consisting of a serine protease inhibitor, a soluble complement regulator, a therapeutic antibody or an antigen-binding fragment thereof, a complement component inhibitor, and an anaphylatoxin receptor antagonist. 
     
     
         6 . (canceled) 
     
     
         7 . The method of  claim 1 , wherein the mutation in the DNA sequence of the CD55 gene or in the RNA sequence of the CD55 transcript results in a near to complete absence of CD55 protein expression or the expression of CD55 protein with substantially diminished function or that is non-functional. 
     
     
         8 . The method of  claim 1 , wherein the mutation in the DNA sequence of the CD55 gene is at least one of:
 c.149-150delAA;   c.149-150insCCTT;   c.109delC;   c.800G>C;   c.287-1G>C;   c.149-150delAAinsCCTT; or   combinations thereof.   
     
     
         9 . The method of  claim 1 , wherein detecting includes at least one of:
 sequencing at least a portion of the CD55 gene or the CD55 transcript or the CD55 cDNA;   or   contacting a labeled nucleic acid probe to at least a portion of the CD55 gene or the CD55 transcript or the CD55 cDNA; or   contacting at least a portion of the CD55 gene or CD55 transcripts or cDNA thereof with a microarray; or   combinations thereof.   
     
     
         10 . The method of  claim 9 , wherein hybridization of the labeled nucleic acid probe is indicative of a mutation in the portion of the CD55 gene or CD55 transcript. 
     
     
         11 . The method of  claim 9 , wherein hybridization of the labeled nucleic acid probe is indicative of the portion of the CD55 gene or CD55 transcript having a wild-type sequence at the location of hybridization. 
     
     
         12 . The method of  claim 9 , wherein sequencing at least a portion of the CD55 gene or CD55 transcript or cDNA thereof includes amplifying at least one region of interest for sequencing with at least one polymerase chain reaction (PCR) with at least one of the following primer sets: 
       
         
           
                 
                 
               
                     
                   (Exon 1) 
                 
                     
                   CTACTCCACCCGTCTTGTTTGT 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   TTTGGGGGTTAAGGATACAGTC; 
                 
                     
                     
                 
                     
                   (Exon 2) 
                 
                     
                   CAGGTGTGGCATTTCAAGG 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   ACCCTGGGGTTTAGTAACGC; 
                 
                     
                     
                 
                     
                   (Exon 3) 
                 
                     
                   AAGTACTAAATATGCGCAAAGCAG 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   ATGGTCCTATCAAGAAACATCC; 
                 
                     
                     
                 
                     
                   (Exon 4) 
                 
                     
                   GTTACCTTCTTTGTGTGTATGCC 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   GCTGTGAATACCAGTCATGAAAC; 
                 
                     
                     
                 
                     
                   (Exon 5) 
                 
                     
                   AACCTGGAGAATTTGAGGAAAG 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   TGTGCTAATATTCTTAAGGGGC; 
                 
                     
                     
                 
                     
                   (Exon 6) 
                 
                     
                   GCATTTATAAGCATCTCTTGTTGG 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   TCATTGAATGTCTGCAACCC; 
                 
                     
                     
                 
                     
                   (Exon 7) 
                 
                     
                   CTAGGTGTTTGTGGGGAGAGAG 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   TCTGGTGGGTTTCTGAAGAGTT; 
                 
                     
                     
                 
                     
                   (Exon 8) 
                 
                     
                   TTTACGCAGAGTCCTTCAGC 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   CCATTTAATCCTGCAATCTTGG; 
                 
                     
                     
                 
                     
                   (Exon 9) 
                 
                     
                   TGGAAATTTGAGTTGCTTTCG 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   TCTCCCAGGAATATGGATTG; 
                 
                     
                     
                 
                     
                   (Exon 10)  
                 
                     
                   GCACCCCAAATTAACTGATTC 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   ATGTGATTCCAGGACTGCC; 
                 
                     
                   or combinations thereof. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         13 . The method of  claim 9 , wherein contacting a labeled nucleic acid probe to at least a portion of the CD55 gene or the CD55 transcript or the CD55 cDNA is performed using real-time PCR. 
     
     
         14 . The method of  claim 9 , wherein contacting a labeled nucleic acid probe to at least a portion of the CD55 transcripts or cDNA thereof comprises:
 isolating CD55 transcripts;   reverse transcribing at least a portion of the CD55 transcripts; and   contacting the cDNA with the labeled nucleic acid probe.   
     
     
         15 . The method of  claim 1 , wherein the microarray includes probes designed to detect DNA, transcript, or cDNA mutations that result in the complete absence in CD55 protein or a non-functional CD55 protein. 
     
     
         16 . The method of  claim 9 , wherein detecting CD55 protein comprises: contacting the sample with at least one CD55 binding polypeptide, wherein the binding peptide includes a detectable label, or is an anti-CD55 antibody or a CD55-binding fragment of an antibody. 
     
     
         17 .- 23 . (canceled) 
     
     
         24 . The method of  claim 1 , wherein the detecting of CD55 binding function includes examining the at least one of C3b affinity, C3b avidity, C4b affinity, C4b avidity, or combinations thereof. 
     
     
         25 . The method of  claim 1 , wherein detecting complement deposition includes detecting C3d deposition. 
     
     
         26 . A method of treating a patient having CD55 deficiency, hyperactivation of complement, angiopathic thrombosis and protein losing enteropathy (CHAPLE) or preventing at least one symptom of CHAPLE in a patient at risk of developing the same, the method comprising:
 administering an effective amount of a composition comprising at least one complement inhibitor to a subject in need thereof, wherein the composition is effective in treating or preventing at least one symptom of CHAPLE.   
     
     
         27 . The method of  claim 26 , wherein the complement inhibitor is selected from the group consisting of a serine protease inhibitor, a soluble complement regulator, a therapeutic antibody or an antigen-binding fragment thereof, a complement component inhibitor, and an anaphylatoxin receptor antagonist. 
     
     
         28 . The method of  claim 27 , wherein:
 the serine protease inhibitor is at least one of a C3 convertase inhibitor, a C5 convertase inhibitor, a C1 inhibitor, a C1r inhibitor, a C1s inhibitor, a C2a inhibitor, a MASP-1 inhibitor, a MASP-2 inhibitor, a factor D inhibitor, a factor B inhibitor, a factor I inhibitor or combinations thereof;   the soluble complement regulator is at least one of a soluble form of a membrane cofactor protein (MCP or CD46), a soluble form of a decay-accelerating factor (DAF or CD55), a soluble form of a membrane attack complex-inhibitor protein (MAC-IP or CD59), a soluble form of complement receptor 1 (CD35) or combinations thereof;   the therapeutic antibody or the antigen-binding fragment thereof is at least one polypeptide that binds C3, C3a, C3b, C3 convertase, C5, C5a, C5b, C5 convertase, C7, C8, or C9, factor B, factor D, C4, C2, C1, properdin, a functional blocking antibody of an anaphylatoxin or combinations thereof, wherein said binding inhibits complement activation by at least one of blocking association/binding with other complement proteins, blocking association/binding with receptor proteins, blocking serine protease activity or combinations thereof;   the complement component inhibitor is a peptide, nucleic acids, a synthetic molecule or a combination thereof that disrupts protein functions by steric hindrance or the induction of conformational changes; and   the anaphylatoxin receptor antagonist is at least one of a C5aR antagonist, a C5L2 antagonist, a C3a receptor antagonist, a functional blocking antibody of an anaphylatoxin or combinations thereof.   
     
     
         29 .- 37 . (canceled) 
     
     
         38 . A therapeutic composition for treating or preventing at least one symptom of CD55 deficiency, hyperactivation of complement, angiopathic thrombosis and protein losing enteropathy (CHAPLE), the composition comprising:
 an effective amount of two or more agents, wherein at least one of the agents is a complement inhibitor to a subject in need thereof; and   a pharmaceutically acceptable carrier,   wherein the composition is effective in treating or preventing at least one symptom of CHAPLE.   
     
     
         39 . The composition of  claim 38 , wherein the complement inhibitor is selected from the group consisting of a serine protease inhibitor, a soluble complement regulator, a therapeutic antibody or an antigen-binding fragment thereof, a complement component inhibitor, and an anaphylatoxin receptor antagonist. 
     
     
         40 . (canceled) 
     
     
         41 . The composition of  claim 39 , wherein:
 the serine protease inhibitors is at least one of a C3 convertase inhibitor, a C5 convertase inhibitor, a C1 inhibitor, a C1r inhibitor, a C1s inhibitor, a C2a inhibitor, a MASP-1 inhibitor, a MASP-2 inhibitor, a factor D inhibitor, a factor B inhibitor, a factor I inhibitor or combinations thereof;   the soluble complement regulator is at least one of a soluble form of a membrane cofactor protein (MCP or CD46), a soluble form of a decay-accelerating factor (DAF or CD55), a soluble form of a membrane attack complex-inhibitor protein (MAC-IP or CD59), a soluble form of complement receptor 1 (CD35) or combinations thereof;   the therapeutic antibody or the antigen-binding fragment thereof is at least one polypeptide that binds C3, C3a, C3b, C3 convertase, C5, C5a, C5b, C5 convertase, C7, C8, or C9, factor B, factor D, C4, C2, C1, properdin, a functional blocking antibody of an anaphylatoxin or combinations thereof, wherein said binding inhibits complement activation by at least one of blocking association/binding with other complement proteins, blocking association/binding with receptor proteins, blocking serine protease activity or combinations thereof;   the complement component inhibitor is a peptide, nucleic acids, a synthetic molecule or a combination thereof that disrupts protein functions by steric hindrance or the induction of conformational changes; and   the anaphylatoxin receptor antagonist is at least one of a C5aR antagonist, a C5L2 antagonist, a C3a receptor antagonist, a functional blocking antibody of an anaphylatoxin or combinations thereof.   
     
     
         42 .- 50 . (canceled)