Autosomal recessive limb-girdle muscular dystrophy
-Rare genetic disease
-Rare neurologic disease
Qualitative or quantitative defects of calpain
-Rare genetic disease
Comment:
Limb-girdle muscular dystrophy type 2A is one of the most frequent subtypes of autosomal recessive muscular dystrophy, accounting for up to 30% of all recessive LGMD cases taken together. Pathogenic CAPN3
mutations disrupt multiple homeostatic mechanisms in skeletal muscles, resulting in LGMD2A. Patients carrying two null mutations that result in the premature truncation of protein synthesis are invariably characterized by a higher disease severity than patients with at least one missense mutation. (PMID:27081656).
LGMD2A is an autosomal recessive form of muscular dystrophy primarily affecting the proximal muscles, resulting in difficulty walking. The age at onset varies, but most patients show onset in childhood, and the disorder is progressive. Other features may ... LGMD2A is an autosomal recessive form of muscular dystrophy primarily affecting the proximal muscles, resulting in difficulty walking. The age at onset varies, but most patients show onset in childhood, and the disorder is progressive. Other features may include scapular winging, calf pseudohypertrophy, and contractures (summary by Mercuri et al., 2005). - Genetic Heterogeneity of Autosomal Recessive Limb-Girdle Muscular Dystrophy Autosomal recessive LGMD is a genetically heterogeneous disorder: see LGMD2B (253601) caused by mutation in the dysferlin gene (DYSF; 603009) on 2p13; LGMD2C (253700) caused by mutation in the gamma-sarcoglycan gene (SGCG; 608896) on 13q12; LGMD2D (608099) caused by mutation in the alpha-sarcoglycan gene (SGCA; 600119) on 17q12; LGMD2E (604286) caused by mutation in the beta-sarcoglycan gene (SGCB; 600900) on 4q12; LGDM2F (601287) caused by mutation in the delta-sarcoglycan gene (SGCD; 601411) on 5q33; LGMD2G (601954) caused by mutation in the TCAP gene (604488) on 17q12; LGMD2H (254110) caused by mutation in the TRIM32 gene (602290) on 9q31; LGMD2I (607155) caused by mutation in the FKRP gene (606596) on 19q13; LGMD2J (608807) caused by mutation in the titin gene (TTN; 188840) on 2q24; LGMD2K (609308), caused by mutation in the POMT1 gene (607423) on 9q34; LGMD2L (611307), caused by mutation in the ANO5 gene (608662) on 11p14; LGMD2M (611588), caused by mutation in the FKTN gene (607440) on 9q31; LGMD2N (613158), caused by mutation in the POMT2 gene (607439) on 14q24; LGMD2O (613157), caused by mutation in the POMGNT1 gene (606822) on 1p34; LGMD2Q (613723), caused by mutation in the PLEC1 gene (601282) on 8q24; LGMD2R (615325), caused by mutation in the DES gene (125660) on 2q35-q36; and LGM2S (615356), caused by mutation in the TRAPPC11 gene on 4q35. For a discussion of autosomal dominant LGMD (LGMD1), see 159000. Nigro (2003) provided a review of the molecular bases of autosomal recessive LGMD. Daniele et al. (2007) provided a review of therapeutic strategies in various forms of LGMD, including ongoing studies in gene therapy.
Among 58 patients with LGMD2A confirmed by mutation analysis, Fanin et al. (2004) found that 46 (80%) had a variable degree of calpain-3 protein deficiency determined by immunoblot analysis, and 12 (20%) had normal amounts of calpain-3. The ... Among 58 patients with LGMD2A confirmed by mutation analysis, Fanin et al. (2004) found that 46 (80%) had a variable degree of calpain-3 protein deficiency determined by immunoblot analysis, and 12 (20%) had normal amounts of calpain-3. The probability of having LGMD2A was very high (84%) when patients had a complete calpain-3 deficiency and progressively decreased with increasing amounts of protein detected. CAPN3 gene mutations were identified in 46 of 69 (67%) patients with calpain-3 protein deficiency and in 12 of 139 (9%) patients with normal calpain-3 protein. Patients with severe, early-onset disease usually had no calpain-3 protein, but absent or markedly reduced protein levels were also detected in patients with adult onset. However, almost all patients with normal calpain-3 levels had late or adult onset of the disorder. The diagnosis of calpainopathy is obtained by identifying calpain-3 protein deficiency or mutations in the CALPN3 gene. However, in many patients with LGMD2A, loss-of-function mutations cause enzymatic inactivation of calpain-3 while protein quantity remains normal. The identification of such patients is difficult unless a functional test suggests pursuing a search for mutations. Fanin et al. (2007) used a functional in vitro assay to test calpain-3 autolytic function in a large series of muscle biopsy specimens from patients with unclassified LGMD/hyperCKemia who had been shown to have normal calpain-3 protein quantity. Of 148 muscle biopsy specimens tested, 17 (11%) had lost normal autolytic function. The CAPN3 gene mutations were identified in 15 of the 17 patients (88%), who accounted for about 20% of the total patients with LGMD2A diagnosed in their series. Blazquez et al. (2008) performed a retrospective diagnostic study of CAPN3 mRNA expression in peripheral blood of 26 unrelated patients with LGMD2A, including 14 with known biallelic CAPN3 mutations and 12 with only 1 CAPN3 mutation identified through DNA analysis. The results of peripheral blood mRNA analysis confirmed the known mutations. In addition, 7 (25%) of 28 mutations identified by analyzing white blood cell mRNA were splice site mutations that modified the CAPN3 transcript, but would not have been detected by direct DNA sequencing of coding regions. However, 4 different mRNA transcripts were identified in white blood cells, compared to only 1 known to be expressed in skeletal muscle tissue. These different isoforms were produced by alternative splicing of exons 6, 15, and 16 of the CAPN3 gene. Blazquez et al. (2008) concluded that while analysis of CAPN3 mutations at the mRNA level in peripheral blood is useful, the diagnosis must be confirmed by DNA studies, since the results could be discordant mainly because of nonsense-mediated decay of truncated transcripts. By molecular screening, Fanin et al. (2009) identified 66 different CAPN3 mutations in 94 of 519 patients with LGMD. Of those with mutations, 73% had a quantitative protein defect, 16% had a functional protein defect, and 11% had normal protein quantity. Conversely, CAPN3 mutations were found in 80% of patients with a quantitative defect on western blot analysis and in 88% of patients with a functional defect of calpain-3 on skeletal muscle biopsy. In addition, CAPN3 mutations were found in 10 (5.6%) of 178 patients with normal CAPN3 quantity. Fanin et al. (2009) concluded that systematic investigation for LGMD2A in patients should involve biochemical assays and muscle biopsy evaluation to determine which patients are suitable for genetic analysis. - Differential Diagnosis Walton and Nattrass (1954) introduced the term 'limb-girdle muscular dystrophy' as part of a classification that achieved wide acceptance and did much to resolve earlier confusion. Over the next 30 years it became clear that many inherited and acquired disorders could produce a similar clinical picture, such as nemaline myopathy (e.g., 256030), central core disease (117000), thyrotoxic myopathy, various scapuloperoneal syndromes (e.g., 181430), chronic polymyositis, and, spinal muscular atrophy (e.g., 253300). Many of the early reported cases of 'limb-girdle' muscular dystrophy likely had one of these conditions. Attempting total ascertainment of LGMD in the Lothian area of Scotland, Yates and Emery (1985) collected 10 index cases of adult-onset (at or after age 18) LGMD. In the 10 sibships, only 1 had a second case; however, in this family the 2 brothers may have had Becker muscular dystrophy (BMD; 300376). Assuming recessive inheritance, there was a significant deficiency of affected persons and a great preponderance of males (9 out of 10). Arikawa et al. (1991) analyzed diagnostic muscle biopsies in 41 cases with the clinical diagnosis of limb-girdle muscular dystrophy at the National Institute of Neuroscience in Tokyo. Using immunofluorescence, immunoblot analysis, and PCR analysis to examine diagnostic muscle biopsies, they identified 5 male patients with an abnormal dystrophin (300377) pattern diagnostic of Becker muscular dystrophy and 2 female patients with dystrophin patterns consistent with a manifesting carrier of Duchenne muscular dystrophy (DMD; 310200). Thus, 17% of the limb-girdle patients showed a 'dystrophinopathy,' with 31% (4/13) of isolated males misclassified and 13% (2/15) of isolated females misclassified. The study emphasized the clinical overlap between LGMD and dystrophinopathies and reinforced the necessity of dystrophin protein and gene studies for accurate clinical diagnosis.
In a large study of patients with different forms of muscular dystrophy, Chung and Morton (1959) delineated the common features of limb-girdle muscular dystrophy. Onset usually occurred in childhood, but sometimes in maturity or middle age. Involvement was ... In a large study of patients with different forms of muscular dystrophy, Chung and Morton (1959) delineated the common features of limb-girdle muscular dystrophy. Onset usually occurred in childhood, but sometimes in maturity or middle age. Involvement was first evident in either the pelvic or, less frequently, the shoulder girdle, often with asymmetry of wasting when the upper limbs were first involved. Spread from the lower to the upper limbs or vice versa occurred within 20 years. Pseudohypertrophy of the calves was uncommon, but may have been counterfeited by a stocky build or wasting of the vasti. The rate of progression was variable, but severe disability with inability to walk was seen within 20 to 30 years of onset. Contractures and facial weakness occurred in some in late stages. Age at death was variable with the largest number of patients dying in middle age. In the analysis of Chung and Morton (1959), 59% of cases of limb-girdle muscular dystrophy could be ascribed to autosomal recessive inheritance; the remainder were sporadic cases of unknown etiology. Richard et al. (1999) reviewed the clinical information on 163 LGMD2A patients with calpain-3 mutations. They noted that Fardeau et al. (1996) had defined precise clinical features, first noted in patients from Reunion Island. LGMD2A was characterized mainly by a symmetric, very selective atrophic involvement of limb-girdle and trunk muscles, with the gluteus maximus and thigh adductors being most affected. The same pattern of muscle involvement was also reported for three-fourths of the examined metropolitan French patients, with occasionally minor variations around this pattern. Similar findings were found in LGMD2A patients of Turkish or Basque origin. Mean age at onset was 13.7 years (range, 2 to 40 years) and the mean age at loss of walking ability was 17.3 years (range, 5 to 39 years). No sex difference was evident in age at onset or disease progression. Using linkage studies to characterize 13 Brazilian families with autosomal recessive LGMD, Passos-Bueno et al. (1996) found that the approximately 33% of families had LGMD2A and 33% had LGMD2B, whereas 17% had LGMD2D, and less than 10% had LGMD2C. Patients with LGMD2B appeared to have the mildest phenotype, with an average age at onset that was significantly later than for patients with LGMD2A. Passos-Bueno et al. (1999) studied 140 patients from 40 Brazilian families with one of 7 autosomal recessive limb-girdle muscular dystrophies. All LGMD2E and LGMD2F patients had a severe phenotype; considerable inter- and intrafamilial variability was observed in all other types of LGMD. Comparison between 40 LGMD2A patients and 52 LGMD2B patients showed that LGMD2A patients had a more severe course and higher frequency of calf hypertrophy (86% vs 13%), and that LGMD2B patients were more likely to be unable to walk on toes (70% vs 18%). Mercuri et al. (2005) reported skeletal muscle MRI findings of 7 patients with LGMD2A who had early contractures. The 3 younger patients were able to walk independently and had selective impairment of the adductor magnus and semimembranosus muscles. The 4 older patients all had restricted ambulation and had similar involvement of the adductor magnus as well as more diffuse involvement of the posterolateral thigh muscles and vastus intermedius. Two 'control' patients with LGMD2A without contractures showed similar muscle involvement. Mercuri et al. (2005) suggested that patients with LGMD2A have a somewhat unique pattern of muscle involvement on MRI, which may serve to differentiate the disorder from Emery-Dreifuss muscular dystrophy (EDMD; see, e.g., 310300) and Bethlem myopathy (158810), both of which have phenotypic similarities to LGMD2A. LGMD2A is characterized by a wide variability in clinical features and rate of progression. Patients with 2 null mutations usually have a rapid course, but in the remaining cases (2 missense mutations or compound heterozygote mutations) prognosis is uncertain. Fanin et al. (2007) conducted a systematic histopathologic, biochemical, and molecular investigation of 24 LGMD2A patients, subdivided according to rapid or slow disease progression to determine if some parameters could correlate with disease progression. They found that muscle histopathology score and the extent of regenerating and degenerating fibers could be correlated with a rate of disease course. Comparison of clinical and muscle histopathologic data between LGMD2A and 4 other types of LGMD (LGMD2B-E) also gave another significant result. They found that LGMD2A has significantly lower levels of dysmorphic features (i.e., degenerating and regenerating fibers) and higher levels of chronic changes (i.e., lobulated fibers) compared with other LGMDs, particularly LGMD2B (253601). Fanin et al. (2007) concluded that these results might explain the observation that atrophic muscle involvement seems to be a clinical feature peculiar to LGMD2A patients. - Eosinophilic Myositis Krahn et al. (2006) reported 6 unrelated patients originally diagnosed with eosinophilic myositis based on skeletal muscle biopsy in the first decade of life (age at diagnosis 3 to 11 years). All patients presented initially with increased serum creatine kinase. Skeletal muscle biopsies showed focal inflammatory lesions with eosinophilic infiltration and no evidence of parasites; some biopsies showed necrotic fibers. Clinically, 1 patient had only motor fatigability, 2 had mild motor clumsiness, and 1 had difficulty walking on heels. Two of the older patients, ages 16 and 11, respectively, had more significant muscle weakness. Three patients had hypereosinophilia on peripheral blood count. Western blot analysis of 1 patient showed loss of calpain-3. All patients were found to have homozygous or compound heterozygous mutations in the CAPN3 gene (see, e.g., 114240.0006). Krahn et al. (2006) suggested that eosinophilic myositis may be an early and transient feature in calpainopathies, because it was not present in biopsies from older patients with typical LGMD2A. Krahn et al. (2011) reported 5 additional patients with an initial diagnosis of eosinophilic myositis who were found to be homozygous or compound heterozygous for CAPN3 mutations. One adult presented with progressive proximal weakness in her early twenties. The other 4 patients were children who presented in the first decade with increased serum creatine kinase and blood eosinophilia, which was intermittent in 2 cases. In addition, a 5-year-old boy with 2 CAPN3 mutations who presented with proximal muscle weakness of the lower limbs was found to have a focal lymphocytic infiltrate of CD8+ T cells on muscle biopsy. In a retrospective analysis of muscle biopsies from 17 patients with genetically confirmed LGMD2A, Krahn et al. (2011) identified inflammatory changes with presence of eosinophils in 5 patients. The average disease duration at biopsy for patients without or with eosinophils was 13.9 and 6 years, respectively. The findings highlighted eosinophilic infiltration as an early component in primary calpainopathy.
In families with LGMD2 linked to chromosome 15, Richard et al. (1995) identified mutations in the calpain-3 gene; in all, 15 nonsense, splice site, frameshift, or missense calpain-3 mutations were found to segregate with the disease. Six of ... In families with LGMD2 linked to chromosome 15, Richard et al. (1995) identified mutations in the calpain-3 gene; in all, 15 nonsense, splice site, frameshift, or missense calpain-3 mutations were found to segregate with the disease. Six of the mutations were found in Reunion Island patients. Richard et al. (1999), who referred to this disorder as 'calpainopathy,' stated that 97 distinct pathogenic CAPN3 mutations had been identified: 4 nonsense mutations, 32 deletions/insertions, 8 splice site mutations, and 53 missense mutations, together with 12 polymorphisms and 5 unclassified variants. The mutations, most of which represented private variants, were distributed along the entire length of the CAPN3 gene.
Pfaendler (1950) reported an affected Swiss pedigree which was studied further by Touraine (1955). Jackson and Carey (1961) found the same type of autosomal recessive muscular dystrophy in the descendants of Swiss immigrants in an Amish isolate in ... Pfaendler (1950) reported an affected Swiss pedigree which was studied further by Touraine (1955). Jackson and Carey (1961) found the same type of autosomal recessive muscular dystrophy in the descendants of Swiss immigrants in an Amish isolate in Indiana. Moser et al. (1966) found autosomal recessive muscular dystrophy to be 4 times more frequent in the Canton of Berne than in other countries studied. He mapped the places of origin of the parents of cases within the canton, which proved to be the same area as those from which the Amish family names were derived. Urtasun et al. (1998) reported the highest prevalence rate of LGMD described to that time in Guipuzcoa, a small mountainous Basque province in northern Spain: 69 per million. Genetic studies demonstrated that 38 cases corresponded to LGMD2A due to calpain-3 gene mutations. Only 1 patient with alpha-sarcoglycanopathy was found, and in 12 patients the genetic defect was not identified. The particular calpain-3 mutation predominant in Basques (exon 22, 2362AG-to-TCATCT; 114240.0006) had only rarely been found in the rest of the world. The clinical characteristics of the patients with calpain-3 gene mutations were quite homogeneous and different from the other groups, allowing for a precise clinical diagnosis. Disease onset was between ages 8 and 15 years, occurring in the pelvic girdle in most cases, and patients became wheelchair-bound between 11 and 28 years after onset. No pseudohypertrophy of calves or contractures were observed. Canki-Klain et al. (2004) found that 550delA (114240.0009) was the most common mutation among Croatian patients with LGMD2A, with a prevalence of 76% of mutant CAPN3 alleles. The detection of 4 healthy 550delA heterozygous individuals yielded a frequency of 1 in 133 (0.75%) in the general Croatian population. All 4 carriers originated from an island and mountain region near the Adriatic, indicating a probable founder effect. Fanin et al. (2005) determined that the prevalence of LGMD2A in northeastern Italy was 9.47 per million. Two founder mutations in the CAPN3 gene, 550delA and R490Q (114240.0010), were identified. Todorova et al. (2007) identified mutations in the CAPN3 gene in 20 (42%) of 48 unrelated Bulgarian patients with muscular dystrophy. Forty percent of the patients were homozygous for the 500delA mutation, and 70% carried it on at least 1 allele. Van der Kooi et al. (2007) found that LGMD2A was the most common form of LGMD in the Netherlands, accounting for 21% (14 of 67) of all families with LGMD studied. Guglieri et al. (2008) found that LGMD2A was the most common form of LGMD in Italy, present in 28.4% of 155 Italian probands. Duno et al. (2008) found that LGMD2A was uncommon in patients of Danish origin, with an approximately 5- to 6-fold lower prevalence in Denmark compared to other European countries.
Calpainopathy (also known as limb-girdle muscular dystrophy 2A, or LGMD2A) is suspected in individuals with the following:...
Diagnosis
Clinical DiagnosisCalpainopathy (also known as limb-girdle muscular dystrophy 2A, or LGMD2A) is suspected in individuals with the following:Proximal muscle weakness (pelvic and/or shoulder girdle) with early onset (age <12 years), adult onset, or late onset (age >30 years) Atrophy of limb and trunk muscles; possible calf hypertrophy Scapular winging, scoliosis, Achilles tendon contracture, and other joint contractures (including hip, knee, elbow, finger, and spine) Waddling gait; tip-toe walking; difficulty in running, climbing stairs, lifting weights, and getting up from the floor or from a chair Sparing of facial, ocular, tongue, and neck muscles Asymptomatic elevated creatine kinase (CK) concentrations, especially in childhood or adolescenceAbsence of cardiomyopathy and intellectual disabilityFamily history consistent with autosomal recessive inheritance TestingSerum creatine kinase (CK) concentration is always elevated (5-80 times normal) from early infancy on, particularly during the active stage of the disease. Serum CK concentration decreases with disease progression. Muscle CT scan or MRI Wasting of muscles predominantly from the posterior compartment of the limbs, evident on clinical examination, is observed by muscle CT scan [Fardeau et al 1996]. The difference in distribution of muscle involvement between calpainopathy and other limb-girdle muscular dystrophies (LGMDs) (i.e., sarcoglycanopathies and dysferlinopathies) makes CT scan analysis an important element in clinical diagnosis (see Differential Diagnosis).MRI scan confirms this selective involvement [Mercuri et al 2005] and shows in some cases edema-like changes [Fischer et al 2005, Borsato et al 2006, Degardin et al 2010, Stramare et al 2010, Wattjes et al 2010]. Muscle MRI is a useful tool in characterizing the degree of muscle atrophy and in some cases also the severity of the disease.Electromyogram (EMG) pattern is typically myopathic (showing small polyphasic potentials), although a normal EMG can also be observed in presymptomatic individuals. Myotonia and spontaneous discharges are not present. Muscle biopsy Histopathology. A variety of morphologic changes and variable muscle involvement may be observed, irrespective of the age of the individual at the time of biopsy. Most individuals have the typical features of an active dystrophic process (increased fiber size variability, increased fibrosis, regenerating fibers, degenerating and necrotic fibers); others have mild and nonspecific myopathic features (increased central nuclei, fiber splitting, lobulated fibers, type 1 fiber predominance) [Luo et al 2011]. The extent of muscle regeneration is less than is typically observed in other LGMDs [Fanin et al 2007a, Saenz et al 2008, Hauerslev et al 2012]. Eosinophilic myositis is an early and transient feature of calpainopathy, which has been reported in individuals with increased CK levels [Brown & Amato 2006, Krahn et al 2006a, Oflazer et al 2009, Krahn et al 2010] and is not present in muscle from older affected individuals with typical calpainopathy. Wide variability can be observed among individuals who are homozygous for the same missense mutation [Chae et al 2001, Fanin et al 2003]. Calpain-3 immunohistochemical analysis. Absent immunohistochemical reaction was reported in muscle from affected individuals in whom the absent protein had previously been detected by immunoblotting [Kolski et al 2008, Charlton et al 2009]. This method appears however less sensitive than immunoblotting in detecting partial protein reduction. Calpain-3 immunoblot analysis of muscle biopsy is the most useful diagnostic tool in calpainopathy. Approximately 80% of individuals with CAPN3 mutations show variable levels of calpain-3 protein deficiency (~58% of affected individuals have no detectable calpain-3 protein; 22% have partial reduction of the amount of protein) and 20% have a normal amount of protein (but with loss of protein function) [Fanin et al 2004, Groen et al 2007, Fanin et al 2009a]. Note: The results of calpain-3 immunoblot analysis need to be interpreted with caution, as the analysis is neither completely sensitive (i.e., it can yield false negative results) nor completely specific (i.e., it can yield false positive results). In particular, calpain-3 protein levels can be partially reduced in other muscular dystrophies such as dysferlinopathy and Udd muscular dystrophy (titinopathy) [Anderson et al 2000, Fanin et al 2001, Haravuori et al 2001]. Moreover, although calpain-3 protein is extremely stable in human muscle over time [Anderson et al 1998], the amount of protein can be partially reduced by the degradation that occurs when muscle tissue is handled or stored under conditions that promote rapid calpain-3 autolysis (especially partial thawing and exposure to moisture) [Anderson et al 1998, Fanin et al 2003]. The probability that an individual has calpainopathy (LGMD2A) is very high (84%) if calpain immunoblotting testing shows complete calpain-3 deficiency; the probability progressively decreases with the amount of protein detected [Fanin et al 2004, Fanin et al 2009a]. Assay of calpain-3 autolytic function in muscle is better than conventional immunoblot analysis in identifying those affected individuals in whom CAPN3 mutations affect the autolytic function of calpain-3 protein rather than the quantity of calpain-3 protein [Fanin et al 2007b]. Molecular Genetic TestingGene. CAPN3, which encodes proteolytic enzyme calpain-3, is the only gene in which mutations are known to cause calpainopathy. Clinical testing Table 1. Summary of Molecular Genetic Testing Used in CalpainopathyView in own windowGene SymbolTest MethodMutations DetectedMutation Detection Frequency by Test Method 1Test AvailabilityCAPN3Sequence analysis of genomic or cDNA 2Sequence variants 3~99% 4Clinical
Deletion / duplication analysis 5Partial- and whole-gene deletions / duplicationsUnknown 61. The ability of the test method used to detect a mutation that is present in the indicated gene2. Sequence analysis of cDNA (from muscle or blood) may be more efficient than exon-by-exon genomic sequencing [Richard & Beckmann 1995]; functional consequences of variants detected in non-coding regions and deep-intronic mutations have been reported [Krahn et al 2007, Blazquez et al 2008, Nascimbeni et al 2010]. 3. Examples of mutations detected by sequence analysis may include small intragenic deletions/insertions and missense, nonsense, and splice site mutations.4. Although CAPN3 mutations are distributed throughout the length of the gene [Richard et al 1999], in some populations most mutant alleles are clustered in a limited numbers of exons [Anderson et al 1998, De Paula et al 2002, Zatz et al 2003, Fanin et al 2004, Piluso et al 2005, Leiden Muscular Dystrophy Pages]. (see also Molecular Genetics). Some laboratories serving a specific population may offer targeted mutation analysis of a mutation panel.5. Testing that identifies deletions/duplications not readily detectable by sequence analysis of the coding and flanking intronic regions of genomic DNA; included in the variety of methods that may be used are: quantitative PCR, long-range PCR, multiplex ligation-dependent probe amplification (MLPA), and chromosomal microarray (CMA) that includes this gene/chromosome segment.6. Large genomic mutations [Richard et al 1999] including the frame-shift deletion of exons 2-8 [Todorova et al 2007] are a cause of calpainopathy.Interpretation of test results. For issues to consider in interpretation of sequence analysis results, click here.Information on specific allelic variants may be available in Molecular Genetics (see Table A. Genes and Databases and/or Pathologic allelic variants).Testing StrategyTo confirm/establish the diagnosis in a proband. The testing strategy used in the diagnosis of calpainopathy in a proband differs depending on the availability of diagnostic muscle biopsy. Elements of the diagnostic approach are summarized in Figure 1.FigureFigure 1. Diagnostic algorithm when calpainopathy (LGMD2A) is suspected Individuals for whom muscle biopsy is available. Diagnostic muscle biopsy allows screening by western blot of multiple proteins that are responsible for different forms of LGMD. After exclusion of other protein defects (i.e., sarcoglycans, dystrophin, dysferlin, caveolin), calpain-3 protein immunoblot remains the most useful strategy in the diagnosis of calpainopathy [Pollitt et al 2001, Fanin et al 2009b]. The subsequent steps, aimed at identifying gene mutations, vary according to the findings obtained from protein analysis: In individuals with immunoblot-confirmed calpain-3 deficiency, the detection rate for one or two pathogenic CAPN3 mutations is approximately 84%. If two mutant alleles are identified, the genetic diagnosis is confirmed. In individuals with immunoblot-confirmed calpain-3 protein deficiency and no identified CAPN3 mutations, genetic diagnosis of calpainopathy can neither be established nor excluded. In individuals with normal amounts of calpain-3 protein on immunoblot analysis of muscle biopsy tissue, the diagnosis of calpainopathy appears unlikely but cannot be excluded because of possible functional enzyme defects. Such individuals have a residual risk of calpainopathy of approximately 9%. In these individuals, sequencing of CAPN3 can be pursued but the yield will be low. Individuals for whom muscle biopsy is unavailable. Sequence analysis of CAPN3 is the optimal diagnostic step in these individuals [Nigro et al 2011]. Single gene testing. One strategy for molecular diagnosis of a proband suspected of having calpainopathy is CAPN3 sequence analysis, which is expected to identify a mutation in fewer than 40% of individuals with LGMD (of which calpainopathy (LGMD2A) is the most common form) in non-consanguineous populations [Kawai et al 1998, Chou et al 1999, Zatz et al 2000, Bushby & Beckmann 2003, Guglieri et al 2008, Fanin et al 2009b]. Multi-gene panel. Another strategy for molecular diagnosis of a proband suspected of having calpainopathy is use of a multi-gene panel. Note: The genes included and the methods used in multi-gene panels vary by laboratory and over time; a panel may not include a specific gene of interest.Carrier testing for at-risk relatives requires prior identification of the disease-causing mutations in the family. Note: Carriers are heterozygotes and are not at risk of developing the disorder.Prenatal diagnosis for at-risk pregnancies requires prior identification of the disease-causing mutations in the family. Genetically Related (Allelic) DisordersNo other phenotypes are associated with mutations in CAPN3.
Calpainopathy is characterized by symmetric and progressive weakness of proximal (limb-girdle) muscles. The age at onset of muscle weakness ranges from two to 40 years. Early motor milestones are usually normal. Intra- and interfamilial clinical variability ranges from severe to mild [Richard et al 1999]. ...
Natural History
Calpainopathy is characterized by symmetric and progressive weakness of proximal (limb-girdle) muscles. The age at onset of muscle weakness ranges from two to 40 years. Early motor milestones are usually normal. Intra- and interfamilial clinical variability ranges from severe to mild [Richard et al 1999]. Three calpainopathy phenotypes have been identified based on the distribution of muscle weakness and age at onset:Pelvifemoral LGMD (Leyden-Möbius) phenotype, the most frequently observed calpainopathy phenotype. Muscle weakness is first evident in the pelvic girdle and later in the shoulder girdle. Onset can be early (age <12 years), adult, or late (age >30 years). Individuals with early onset and rapid disease course usually have pelvifemoral LGMD. Scapulohumeral LGMD (Erb) phenotype. Muscle weakness is first evident in the shoulder girdle and later in the pelvic girdle. Early onset is infrequent; the disease course is variable, but usually milder than that in the pelvifemoral phenotype. HyperCKemia. Asymptomatic individuals have high serum CK concentrations only. HyperCKemia may be considered a presymptomatic stage of calpainopathy, as it is usually observed in children or young individuals [Fanin et al 2009b, Kyriakides et al 2010].The first clinical findings of calpainopathy are usually the tendency to walk on tiptoes, difficulty in running, and scapular winging. Waddling gait and slight hyperlordosis are frequently observed in the early stage of the disease. Symmetric weakness of proximal more than distal muscles is evident in the limbs, trunk, and periscapular area. Marked laxity of the abdominal muscles is frequently seen [Bushby 1999, Pollitt et al 2001]. The gluteus maximus, thigh adductors, and posterior compartment of the limbs are most affected [Fardeau et al 1996, Dincer et al 1997, Topaloglu et al 1997, Urtasun et al 1998]. Early Achilles tendon shortening and scoliosis may be present.Some individuals report muscle pain and exercise intolerance [Penisson-Besnier et al 1998]. In some individuals, eosinophilic myositis was also found [Krahn et al 2006b]. Calf hypertrophy is rarely observed, and in a number of cases, significant atrophy is observed. Facial and neck muscles are usually spared. Macroglossia has not been described.In the advanced stage of the disease, the inability to climb stairs, to rise up from a chair, or to get up from the floor is common. Joint contractures (in the hips, knees, elbows, and fingers) are common; rigid spine is occasionally observed [Pollitt et al 2001]. Foot drop may occasionally be present [Burke et al 2010]. Respiratory insufficiency with reduced lung vital capacity may be present. Cardiomyopathy is uncommon. Intelligence is normal. The asymptomatic stage may be relatively long in some affected individuals, especially in females. The disease is invariably progressive and loss of ambulation occurs approximately ten to 30 years after the onset of symptoms (between ages 10 and 48 years) [Richard et al 1999, Zatz et al 2003, Saenz et al 2005, Angelini et al 2010].
Genotype-phenotype correlation in calpainopathy is complex and often complicated by the fact that most individuals are compound heterozygotes for CAPN3 mutations. ...
Genotype-Phenotype Correlations
Genotype-phenotype correlation in calpainopathy is complex and often complicated by the fact that most individuals are compound heterozygotes for CAPN3 mutations. Null homozygous mutations are generally associated with a severe phenotype. Although it has been suggested that homozygous missense mutations are usually associated with a milder phenotype than null mutations [Fardeau et al 1996, Anderson et al 1998, Richard et al 1999, Chae et al 2001], the phenotypic consequences of homozygous missense mutations are more difficult to predict [Richard et al 1997, Bushby 1999, De Paula et al 2002, Saenz et al 2011]. Wide clinical variability has been described among individuals homozygous for the same missense mutation, even among individuals within the same family [Fardeau et al 1996, Kawai et al 1998, Penisson-Besnier et al 1998, Richard et al 1999, Fanin et al 2004, Saenz et al 2005, Schessl et al 2008].No direct correlations have been observed between the severity of the phenotype and the amount of calpain-3 protein detected by calpain-3 immunoblot analysis [Anderson et al 1998, Zatz et al 2003, Gallardo et al 2011]. Affected individuals with either no detectable protein or normal amounts of protein have varying severity of the clinical phenotype [De Paula et al 2002, Fanin et al 2004]. Whereas null mutations are usually associated with a lack of detectable protein, missense mutations have variable and unpredictable consequences, which depend at least partially on the quantity of protein.
Other forms of autosomal recessive limb-girdle muscular dystrophy (LGMD2) (i.e., LGMD2B – LGMD2L) cannot be distinguished from calpainopathy on clinical grounds, although calpainopathy generally has a later onset and is relatively mild, particularly by comparison with sarcoglycanopathies. Immunoblot analysis of muscle biopsy for candidate proteins (sarcoglycans, dysferlin, telethonin, titin) can help establish the correct diagnosis. (See Limb-Girdle Muscular Dystrophy Overview.) ...
Differential Diagnosis
Other forms of autosomal recessive limb-girdle muscular dystrophy (LGMD2) (i.e., LGMD2B – LGMD2L) cannot be distinguished from calpainopathy on clinical grounds, although calpainopathy generally has a later onset and is relatively mild, particularly by comparison with sarcoglycanopathies. Immunoblot analysis of muscle biopsy for candidate proteins (sarcoglycans, dysferlin, telethonin, titin) can help establish the correct diagnosis. (See Limb-Girdle Muscular Dystrophy Overview.) Facioscapulohumeral muscular dystrophy (FSHD) shares some clinical and laboratory features with Erb muscular dystrophy (one of the calpainopathy phenotypes): muscle weakness with onset in the shoulder girdle, scapular winging, elevated serum CK concentration, and nonspecific myopathic changes on muscle biopsy. Facial muscle weakness and asymmetric muscle involvement, which can be observed in FSHD, are uncommon in calpainopathy. Inheritance is autosomal dominant. Becker muscular dystrophy (BMD) should be considered in males with:Onset of weakness in the lower girdle muscles in adolescence or adulthood; and Elevated serum CK concentrations; andOne of the following: X-linked recessive pattern of inheritance (sometimes only resulting in elevated serum CK concentration in the proband's mother); No family history of muscle disease;Heart involvement (mainly dilated cardiomyopathy)Diagnosis can be established by dystrophin immunoblot analysis on muscle biopsy or molecular genetic testing of DMD. (See Dystrophinopathies.) Metabolic myopathy. Calpainopathy has been reported in individuals with asthenia, myalgias, exercise intolerance, lower-limb proximal muscle weakness, and excessive lactate production after aerobic exercise [Penisson-Besnier et al 1998]. Such individuals may be difficult to diagnose. Myopathy with contractures. The phenotype of calpainopathy may include muscle weakness with severe tendon contractures [Pollitt et al 2001], raising the possibility of Emery-Dreifuss muscular dystrophy. Note to clinicians: For a patient-specific ‘simultaneous consult’ related to this disorder, go to , an interactive diagnostic decision support software tool that provides differential diagnoses based on patient findings (registration or institutional access required).
To establish the extent of disease in an individual diagnosed with calpainopathy, a complete physical evaluation including the grading of muscle strength in single muscles and the analysis of several functional performances is recommended. ...
Management
Evaluations Following Initial DiagnosisTo establish the extent of disease in an individual diagnosed with calpainopathy, a complete physical evaluation including the grading of muscle strength in single muscles and the analysis of several functional performances is recommended. Treatment of ManifestationsAppropriate management, tailored to each individual, can improve quality of life and prolong survival. The general approach is based on the typical progression and complications of individuals with LGMD as described by McDonald et al [1995], Bushby [1999], and Norwood et al [2007].Physical therapy and stretching exercises can promote mobility, prolong walking, slow the disease progression, in particular by maintaining joint flexibility. Technical aids can also compensate for the loss of certain motor abilities; canes, walkers, orthotics, and wheelchairs enable individuals to regain independence. Surgical intervention may be required for correction of orthopedic complications including foot deformities, scoliosis, and Achilles tendon contractures. Occasionally, scapular fixation may be required for particularly problematic scapular winging.In the late stage of the disease, chronic respiratory insufficiency may occur and the use of respiratory aids may be indicated to prolong survival. Affected individuals should be monitored for signs of hypoventilation and for chest infections, to which they have an increased susceptibility.Social and emotional support help to improve the quality of life, to maximize a sense of social involvement and productivity, and to reduce the sense of social isolation [Eggers & Zatz 1998]. Prevention of Secondary ManifestationsThere are a number of measures that reverse disease manifestations in a symptomatic person, i.e. control of weight gain, prevention of joint contractures by means of physical therapy and stretching exercises, and in the advanced stage, control of respiratory insufficiency. Physical therapy and stretching exercises can help to slow disease progression; therefore, a physical therapy program should be instituted early after diagnosis.SurveillanceAnnual monitoring of muscle strength, joint range of motion, and respiratory function is recommended.Monitoring for orthopedic complications, such as foot deformities, scoliosis, and Achilles tendon contractures, is recommended.Agents/Circumstances to AvoidStrenuous and eccentric muscle exercise should be discouraged as it exacerbates muscle necrosis and could precipitate the onset of weakness or accelerate muscle wasting.Body weight should be controlled to avoid obesity as well as excessive weight loss? (atrophy of muscles can be accelerated by loss of muscle proteins).Physical trauma, bone fractures, and immobility can induce disuse atrophy and thus should be avoided.Although no association of the disease with malignant hyperthermia is reported, the use of succinylcholine and halogenated anesthetic agents should be avoided when possible. (See Malignant Hyperthermia Susceptibility.)While the specific mechanism whereby cholesterol-lowering agents (e.g., statins) may produce muscle lesions is unknown, such drugs should be avoided when possible.Evaluation of Relatives at RiskRelatives at risk (e.g., sibs of probands) should be clinically examined, especially if they may be affected. Molecular genetic testing can be offered to at-risk relatives when clinical findings suggest calpainopathy and the disease-causing mutations in the family are known. See Genetic Counseling for issues related to testing of at-risk relatives for genetic counseling purposes.Pregnancy ManagementWomen with calpainopathy do not have impaired uterine smooth muscle strength or function and typically have uncomplicated pregnancies. Therapies Under InvestigationThe safety and efficacy of AAV-mediated calpain-3 gene transfer in a mouse model of calpainopthy has been reported [Bartoli et al 2006].Search ClinicalTrials.gov for access to information on clinical studies for a wide range of diseases and conditions.
Information in the Molecular Genetics and OMIM tables may differ from that elsewhere in the GeneReview: tables may contain more recent information. —ED....
Molecular Genetics
Information in the Molecular Genetics and OMIM tables may differ from that elsewhere in the GeneReview: tables may contain more recent information. —ED.Table A. Calpainopathy: Genes and DatabasesView in own windowLocus NameGene SymbolChromosomal LocusProtein NameLocus SpecificHGMDLGMD2A
CAPN315q15.1Calpain-3CAPN3 homepage - Leiden Muscular Dystrophy pagesCAPN3Data are compiled from the following standard references: gene symbol from HGNC; chromosomal locus, locus name, critical region, complementation group from OMIM; protein name from UniProt. For a description of databases (Locus Specific, HGMD) to which links are provided, click here.Table B. OMIM Entries for Calpainopathy (View All in OMIM) View in own window 114240CALPAIN 3; CAPN3 253600MUSCULAR DYSTROPHY, LIMB-GIRDLE, TYPE 2A; LGMD2ANormal allelic variants. The longest CAPN3 transcript variant comprises 24 exons and covers a genomic region of 50 kb. It is expressed as a 3.5-kb transcript (2466 coding nucleotides). CAPN3 encodes a number of alternatively spliced transcripts [Herasse et al 1999]. Alternate promoters and alternative splicing result in multiple transcript variants encoding different isoforms and some variants are ubiquitously expressed [De Tullio et al 2003, Kawabata et al 2003].Pathologic allelic variants. More than 350 disease-causing mutations have been reported, most of which are private mutations. They are distributed throughout CAPN3, although a few exons are more frequently involved [Anderson et al 1998, De Paula et al 2002, Zatz et al 2003, Fanin et al 2004] (see Molecular Genetic Testing). Most mutations are single-nucleotide changes. Approximately 70% of mutant alleles have missense mutations; the remaining are a variety of null mutations (small deletions or insertions causing frameshift and premature stop codon, nonsense, and splice site mutations) [Richard et al 1999]. Large genomic mutations [Richard et al 1999, Todorova et al 2007], synonymous codon mutations [Richard & Beckmann 1995], and intronic mutations causing aberrant splicing are further causes of calpainopathy [Krahn et al 2007, Blazquez et al 2008, Nascimbeni et al 2010]. In some populations most mutant alleles are clustered in a limited numbers of exons [Anderson et al 1998, De Paula et al 2002, Zatz et al 2003, Fanin et al 2004, Piluso et al 2005, Leiden Muscular Dystrophy Pages]. Approximately 80% of mutations reported in Brazil are clustered in exons 1, 2, 4, 5, 11, and 22 only [Zatz et al 2003]. Approximately 87% of mutations reported in Italy are found in exons 1, 4, 5, 8, 10, 11, and 21 [Fanin et al 2004]. Approximately 80% of mutations reported in France are clustered in exons 1, 4, 7, 10, 11, 13, 19, and 22 [Krahn et al 2006a].Many mutations have been observed repeatedly in different populations; the c.550delA mutation is the most common allele among individuals from different European countries [Richard et al 1999]. Single mutations recur in the following populations, most likely the result of a founder effect: Reunion Island (c.946-1G>A mutation) [Fardeau et al 1996] Old Order Amish in northern Indiana (c.2306>A mutation) [Young et al 1992]Russia, Croatia, Turkey, Czech Republic, Bulgaria, Germany (c.550delA mutation) [Dincer et al 1997, Pogoda et al 2000, Canki-Klain et al 2004, Chrobakova et al 2004, Balci et al 2006, Hanisch et al 2007, Todorova et al 2007] Basque region of Spain; Brazil (c.2362_2363delAGinsTCATCT mutation) [Urtasun et al 1998, De Paula et al 2002] Japan (c.1795-1796insA mutation) [Kawai et al 1998, Chae et al 2001] Fersina River valley in the Italian Alps (c.1193+6T>A mutation) [Fanin et al 2012]For more information, see Table A.Table 2. Selected CAPN3 Pathologic Allelic VariantsView in own windowDNA Nucleotide Change (Alias 1) Protein Amino Acid ChangeReference Sequence c.550delA p.Thr184Argfs*33NM_000070.2 NP_000061.1c.946-1G>A (IVS6-1G>A)-- c.1193+6T>A--c.1795_1796insA p.Thr599Asnfs*30c.2306G>A p.Arg769Gln c.2362_2363delAGinsTCATCT (2362AG>TCATCT) p.Arg788Serfs*13See Quick Reference for an explanation of nomenclature. GeneReviews follows the standard naming conventions of the Human Genome Variation Society (www.hgvs.org). 1. Variant designation that does not conform to current naming conventionsNormal gene product. Calpain-3 is an enzymatic protein of approximately 94 kd molecular weight (also called p94) composed of 821 amino acids. It is the muscle-specific member of a family of Ca++-activated neutral proteases, which cleave proteins into short polypeptides. Calpain-3 is a multidomain protein with three exclusive sequence inserts (NS, IS1, IS2); domain I has a regulatory role, domain II is the proteolytic module, domain III has a C2-like domain, and domain IV binds Ca++ ions [Ono et al 1998]. Calpain-3 is expressed predominantly in skeletal muscle, where it is localized either in the nucleus or in the cytoplasm (where it binds to the protein titin) [Keira et al 2003]. Although the physiologic role of calpain-3 is still under investigation, it is thought to process proteins involved in signaling pathways, transcription factors, calcium transport, and cytoskeletal proteins as part of a process called sarcomere remodeling [Baghdiguian et al 1999, Baghdiguian et al 2001, Kramerova et al 2005, Duguez et al 2006, Kramerova et al 2007, Beckmann & Spencer 2008, Benayoun et al 2008, Kramerova et al 2008, Saenz et al 2008, Fanin et al 2009c, Ono et al 2010, Ermolova et al 2011]. Abnormal gene product. CAPN3 mutations have a loss of function effect on translated protein. Most individuals with calpainopathy have complete or partial calpain-3 protein deficiency on muscle biopsy as a result of premature truncating mutations or increased protein instability (when caused by missense mutations). In 10% to 30% of individuals, muscle biopsies have a normal amount of protein [Talim et al 2001, De Paula et al 2002, Fanin et al 2004, Groen et al 2007, Milic et al 2007, Fanin et al 2009a], even though calpain-3 may have lost its autocatalytic activity and may be functionally inactive [Fanin et al 2003, Fanin et al 2007b].