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Syrup Urine Disease (MSUD) is a genetic disorder caused by a deficiency of branched-chain keto acid dehydrogenase (BCKDH), a mitochondrial multienzyme complex responsible for the oxidative decarboxylation of branched-chain keto acids derived from branched-chain amino acids (BCAA), leucine, isoleucine and valine (for review, see: [1]).\nPatients with MSUD, depending on the mutation, show variable degrees of enzyme deficiency leading to several different clinical phenotypes [1]. Approximately 75% of MSUD patients have the classic form of the disease with BCKDH activity in the range of 0–2% of normal [1]. These patients show markedly elevated levels of BCAA in blood and other body fluids [1]. Besides the classic form, there are other variants of the disease. Patients with the intermediate form of the disease show BCKDH activity in the range of 3–30% of normal. In such patients the onset of the disease is delayed, but there are persistently elevated levels of BCAA [1]. Patients with the intermittent form of MSUD show BCKDH activity in the range of 5–20% and during the asymptomatic phase the blood BCAA levels are normal [1]. The overall incidence of MSUD in the general population is 1:185,000 [1], and in certain population groups, such as Mennonites of Pennsylvania, the incidence is estimated to be as high as 1:176 [2].\nThe BCKDH complex, the deficient enzyme in MSUD, consists of three catalytic proteins, a decarboxylase (E1), a dihydrolipoyl transacylase (E2), and a dihydrolipoyl dehydrogenase (E3). The E1 component is a heterotetramer composed of two α and two β subunits [3]. The E1 and E2 components are specific to BCKDH, whereas E3 is also used by pyruvate and α-ketoglutarate dehydrogenase complexes [3] and the glycine cleavage system [4]. BCKDH is also associated with two regulatory proteins, a specific kinase and a phosphatase which regulate the activity of this enzyme through a phosphorylation (inactivation) and dephosphorylation (activation) cycle of the E1 α subunit [5,6]. Mutations in the genes of the E1 and E2 subunits of BCKDH have been described, however, the majority of MSUD mutations identified thus far are in the E2 subunit [1,7]. To date, cases of MSUD have not been associated with defects in the regulatory kinase and phosphatase [1].\nCurrent management of MSUD patients relies on a strict lifelong dietary restriction of protein or BCAA [1,8]. Such a dietary management of the disease, however, is not entirely satisfactory especially in times of metabolic decompensation due to infection, injuries and other stressors. In spite of dietary intervention, there is significant mortality associated with MSUD and there is a high incidence of mental retardation in survivors [9].\nBecause of the central role of the liver in amino acid metabolism and moderate/high levels of BCKDH activity in human liver [10-12], a few cases of MSUD have recently been treated by liver transplantation [13-16]. While the short-term outcome of liver transplantation is encouraging, long-term effects of this approach are not known. However, these patients are now required to take immunosuppressant drugs for the rest of their lives, often with undesirable side effects. Moreover, the cost associated with liver transplantation and the availability of donor livers are additional limiting factors for the practicality of treatment of this disease.\nBecause of the current unsatisfactory options for the treatment of MSUD, there is a need for improved therapies to combat this disease. An obstacle to developing novel treatments for MSUD has been the lack of a suitable animal model to perform necessary preclinical studies. Although a Hereford calf model with MSUD has been described [17-19], this model is neither readily available nor practical to perform preclinical studies. Furthermore, comparison of this animal model with human MSUD has shown some differences in the pathology of the disease [1], making this animal a less desirable model for the human disease.\nRecently, a N-ethyl-N-nitrosourea (ENU)-induced mutant mouse that phenotypically resembles human MSUD has been described [20]. However, the mutation in this model disrupts a splice site in the mitochondrial branched-chain aminotransferase (BCAT) gene, not in BCKDH, the deficient enzyme in MSUD. Because the mutation is not in BCKDH, the validity of this mutant mouse line for modeling human MSUD is questionable.\nThe objective of the present study was to create genetically engineered murine models of MSUD that mimic the pathology of the classic and intermediate variant forms of the human disease. The classic model was created by targeted inactivation of the E2 subunit of BCKDH by homologous recombination in embryonic stem (ES) cells. The model of intermediate MSUD was created by partial transgenic rescue of the E2 gene knockout. This report describes the generation and characterization of these murine models of MSUD. These models will allow for the development of novel treatment approaches, such as gene or stem cell therapies, to ultimately cure MSUD."}
craft-ca-core-ex-dev
{"project":"craft-ca-core-ex-dev","denotations":[{"id":"T3189","span":{"begin":240,"end":250},"obj":"CHEBI:25754"},{"id":"T3190","span":{"begin":264,"end":290},"obj":"CHEBI:22918"},{"id":"T3191","span":{"begin":279,"end":290},"obj":"CHEBI_SO_EXT:amino_acid"},{"id":"T3192","span":{"begin":292,"end":296},"obj":"CHEBI:22918"},{"id":"T3193","span":{"begin":299,"end":306},"obj":"CHEBI_SO_EXT:leucine"},{"id":"T3194","span":{"begin":308,"end":318},"obj":"CHEBI_SO_EXT:isoleucine"},{"id":"T3195","span":{"begin":323,"end":329},"obj":"CHEBI_SO_EXT:valine"},{"id":"T3196","span":{"begin":391,"end":399},"obj":"SO_EXT:sequence_alteration_entity_or_process"},{"id":"T3197","span":{"begin":426,"end":432},"obj":"CHEBI_GO_SO_EXT:enzyme"},{"id":"T3198","span":{"begin":674,"end":678},"obj":"CHEBI:22918"},{"id":"T3199","span":{"begin":682,"end":687},"obj":"UBERON:0000178"},{"id":"T3200","span":{"begin":698,"end":709},"obj":"UBERON:0006314"},{"id":"T3201","span":{"begin":986,"end":990},"obj":"CHEBI:22918"},{"id":"T3202","span":{"begin":1120,"end":1125},"obj":"UBERON:0000178"},{"id":"T3203","span":{"begin":1126,"end":1130},"obj":"CHEBI:22918"},{"id":"T3204","span":{"begin":1357,"end":1370},"obj":"GO:0017086"},{"id":"T3205","span":{"begin":1386,"end":1392},"obj":"CHEBI_GO_SO_EXT:enzyme"},{"id":"T3206","span":{"begin":1420,"end":1429},"obj":"GO_MOP_EXT:catalysis"},{"id":"T3207","span":{"begin":1430,"end":1438},"obj":"CHEBI_PR_EXT:protein"},{"id":"T3208","span":{"begin":1442,"end":1455},"obj":"GO_EXT:0016831"},{"id":"T3209","span":{"begin":1464,"end":1477},"obj":"CHEBI:50488"},{"id":"T3210","span":{"begin":1464,"end":1490},"obj":"GO_PR_EXT:dihydrolipoyllysine_residue_2_methylpropanoyl_transferase"},{"id":"T3211","span":{"begin":1492,"end":1494},"obj":"PR_EXT:000006300"},{"id":"T3212","span":{"begin":1503,"end":1516},"obj":"CHEBI:50488"},{"id":"T3213","span":{"begin":1503,"end":1530},"obj":"GO_PR_EXT:dihydrolipoamide_dehydrogenase"},{"id":"T3214","span":{"begin":1532,"end":1534},"obj":"PR_EXT:000006506"},{"id":"T3215","span":{"begin":1565,"end":1573},"obj":"GO_EXT:tetrameric_macromolecular_complex"},{"id":"T3216","span":{"begin":1590,"end":1591},"obj":"_FRAGMENT"},{"id":"T3217","span":{"begin":1602,"end":1610},"obj":"PR_EXT:000004686"},{"id":"T3218","span":{"begin":1600,"end":1610},"obj":"PR_EXT:000004687"},{"id":"T3219","span":{"begin":1627,"end":1629},"obj":"PR_EXT:000006300"},{"id":"T3220","span":{"begin":1672,"end":1674},"obj":"PR_EXT:000006506"},{"id":"T3221","span":{"begin":1691,"end":1699},"obj":"CHEBI:15361"},{"id":"T3222","span":{"begin":1691,"end":1699},"obj":"_FRAGMENT"},{"id":"T3223","span":{"begin":1720,"end":1733},"obj":"GO_EXT:0004738"},{"id":"T3224","span":{"begin":1720,"end":1743},"obj":"GO:0045254"},{"id":"T3225","span":{"begin":1704,"end":1719},"obj":"CHEBI_EXT:2_oxoglutarate"},{"id":"T3226","span":{"begin":1704,"end":1733},"obj":"GO_EXT:0004591"},{"id":"T3227","span":{"begin":1704,"end":1743},"obj":"GO:0045240"},{"id":"T3228","span":{"begin":1756,"end":1763},"obj":"CHEBI_SO_EXT:glycine"},{"id":"T3229","span":{"begin":1756,"end":1779},"obj":"GO:0005960"},{"id":"T3230","span":{"begin":1764,"end":1772},"obj":"MOP:0000780"},{"id":"T3231","span":{"begin":1819,"end":1829},"obj":"GO:0065007"},{"id":"T3232","span":{"begin":1830,"end":1838},"obj":"CHEBI_PR_EXT:protein"},{"id":"T3233","span":{"begin":1851,"end":1857},"obj":"GO_EXT:0016301"},{"id":"T3234","span":{"begin":1864,"end":1875},"obj":"GO_EXT:0016791"},{"id":"T3235","span":{"begin":1882,"end":1890},"obj":"GO:0065007"},{"id":"T3236","span":{"begin":1912,"end":1918},"obj":"CHEBI_GO_SO_EXT:enzyme"},{"id":"T3237","span":{"begin":1929,"end":1944},"obj":"GO_PATO_EXT:phosphorylation_process"},{"id":"T3238","span":{"begin":1964,"end":1981},"obj":"GO:0016311"},{"id":"T3239","span":{"begin":2008,"end":2020},"obj":"PR_EXT:000004686"},{"id":"T3240","span":{"begin":2028,"end":2037},"obj":"SO_EXT:sequence_alteration_entity_or_process"},{"id":"T3241","span":{"begin":2045,"end":2050},"obj":"SO_EXT:0000704"},{"id":"T3242","span":{"begin":2065,"end":2085},"obj":"PR_EXT:000006300"},{"id":"T3243","span":{"begin":2137,"end":2146},"obj":"SO_EXT:sequence_alteration_entity_or_process"},{"id":"T3244","span":{"begin":2178,"end":2188},"obj":"PR_EXT:000006300"},{"id":"T3245","span":{"begin":2264,"end":2274},"obj":"GO:0065007"},{"id":"T3246","span":{"begin":2275,"end":2281},"obj":"GO_EXT:0016301"},{"id":"T3247","span":{"begin":2286,"end":2297},"obj":"GO_EXT:0016791"},{"id":"T3248","span":{"begin":2390,"end":2397},"obj":"CHEBI_PR_EXT:protein"},{"id":"T3249","span":{"begin":2401,"end":2405},"obj":"CHEBI:22918"},{"id":"T3250","span":{"begin":2516,"end":2525},"obj":"GO:0008152"},{"id":"T3251","span":{"begin":2780,"end":2785},"obj":"UBERON:0002107"},{"id":"T3252","span":{"begin":2789,"end":2799},"obj":"CHEBI_SO_EXT:amino_acid"},{"id":"T3253","span":{"begin":2789,"end":2810},"obj":"GO:0006520"},{"id":"T3254","span":{"begin":2857,"end":2862},"obj":"NCBITaxon:9606"},{"id":"T3255","span":{"begin":2863,"en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Syrup Urine Disease (MSUD) is a genetic disorder caused by a deficiency of branched-chain keto acid dehydrogenase (BCKDH), a mitochondrial multienzyme complex responsible for the oxidative decarboxylation of branched-chain keto acids derived from branched-chain amino acids (BCAA), leucine, isoleucine and valine (for review, see: [1]).\nPatients with MSUD, depending on the mutation, show variable degrees of enzyme deficiency leading to several different clinical phenotypes [1]. Approximately 75% of MSUD patients have the classic form of the disease with BCKDH activity in the range of 0–2% of normal [1]. These patients show markedly elevated levels of BCAA in blood and other body fluids [1]. Besides the classic form, there are other variants of the disease. Patients with the intermediate form of the disease show BCKDH activity in the range of 3–30% of normal. In such patients the onset of the disease is delayed, but there are persistently elevated levels of BCAA [1]. Patients with the intermittent form of MSUD show BCKDH activity in the range of 5–20% and during the asymptomatic phase the blood BCAA levels are normal [1]. The overall incidence of MSUD in the general population is 1:185,000 [1], and in certain population groups, such as Mennonites of Pennsylvania, the incidence is estimated to be as high as 1:176 [2].\nThe BCKDH complex, the deficient enzyme in MSUD, consists of three catalytic proteins, a decarboxylase (E1), a dihydrolipoyl transacylase (E2), and a dihydrolipoyl dehydrogenase (E3). The E1 component is a heterotetramer composed of two α and two β subunits [3]. The E1 and E2 components are specific to BCKDH, whereas E3 is also used by pyruvate and α-ketoglutarate dehydrogenase complexes [3] and the glycine cleavage system [4]. BCKDH is also associated with two regulatory proteins, a specific kinase and a phosphatase which regulate the activity of this enzyme through a phosphorylation (inactivation) and dephosphorylation (activation) cycle of the E1 α subunit [5,6]. Mutations in the genes of the E1 and E2 subunits of BCKDH have been described, however, the majority of MSUD mutations identified thus far are in the E2 subunit [1,7]. To date, cases of MSUD have not been associated with defects in the regulatory kinase and phosphatase [1].\nCurrent management of MSUD patients relies on a strict lifelong dietary restriction of protein or BCAA [1,8]. Such a dietary management of the disease, however, is not entirely satisfactory especially in times of metabolic decompensation due to infection, injuries and other stressors. In spite of dietary intervention, there is significant mortality associated with MSUD and there is a high incidence of mental retardation in survivors [9].\nBecause of the central role of the liver in amino acid metabolism and moderate/high levels of BCKDH activity in human liver [10-12], a few cases of MSUD have recently been treated by liver transplantation [13-16]. While the short-term outcome of liver transplantation is encouraging, long-term effects of this approach are not known. However, these patients are now required to take immunosuppressant drugs for the rest of their lives, often with undesirable side effects. Moreover, the cost associated with liver transplantation and the availability of donor livers are additional limiting factors for the practicality of treatment of this disease.\nBecause of the current unsatisfactory options for the treatment of MSUD, there is a need for improved therapies to combat this disease. An obstacle to developing novel treatments for MSUD has been the lack of a suitable animal model to perform necessary preclinical studies. Although a Hereford calf model with MSUD has been described [17-19], this model is neither readily available nor practical to perform preclinical studies. Furthermore, comparison of this animal model with human MSUD has shown some differences in the pathology of the disease [1], making this animal a less desirable model for the human disease.\nRecently, a N-ethyl-N-nitrosourea (ENU)-induced mutant mouse that phenotypically resembles human MSUD has been described [20]. However, the mutation in this model disrupts a splice site in the mitochondrial branched-chain aminotransferase (BCAT) gene, not in BCKDH, the deficient enzyme in MSUD. Because the mutation is not in BCKDH, the validity of this mutant mouse line for modeling human MSUD is questionable.\nThe objective of the present study was to create genetically engineered murine models of MSUD that mimic the pathology of the classic and intermediate variant forms of the human disease. The classic model was created by targeted inactivation of the E2 subunit of BCKDH by homologous recombination in embryonic stem (ES) cells. The model of intermediate MSUD was created by partial transgenic rescue of the E2 gene knockout. This report describes the generation and characterization of these murine models of MSUD. These models will allow for the development of novel treatment approaches, such as gene or stem cell therapies, to ultimately cure MSUD."}
craft-ca-core-dev
{"project":"craft-ca-core-dev","denotations":[{"id":"T3120","span":{"begin":1532,"end":1534},"obj":"PR:000006506"},{"id":"T3123","span":{"begin":1600,"end":1610},"obj":"PR:000004687"},{"id":"T3124","span":{"begin":1627,"end":1629},"obj":"PR:000006300"},{"id":"T3125","span":{"begin":1672,"end":1674},"obj":"PR:000006506"},{"id":"T3126","span":{"begin":1691,"end":1699},"obj":"CHEBI:15361"},{"id":"T3127","span":{"begin":1691,"end":1699},"obj":"_FRAGMENT"},{"id":"T3128","span":{"begin":1720,"end":1743},"obj":"GO:0045254"},{"id":"T3129","span":{"begin":1704,"end":1743},"obj":"GO:0045240"},{"id":"T3130","span":{"begin":1756,"end":1779},"obj":"GO:0005960"},{"id":"T3131","span":{"begin":1764,"end":1772},"obj":"MOP:0000780"},{"id":"T3132","span":{"begin":1819,"end":1829},"obj":"GO:0065007"},{"id":"T3133","span":{"begin":1882,"end":1890},"obj":"GO:0065007"},{"id":"T3134","span":{"begin":1964,"end":1981},"obj":"GO:0016311"},{"id":"T3135","span":{"begin":2008,"end":2020},"obj":"PR:000004686"},{"id":"T3136","span":{"begin":2045,"end":2050},"obj":"SO:0000704"},{"id":"T3137","span":{"begin":2065,"end":2085},"obj":"PR:000006300"},{"id":"T3138","span":{"begin":2178,"end":2188},"obj":"PR:000006300"},{"id":"T3139","span":{"begin":2264,"end":2274},"obj":"GO:0065007"},{"id":"T3140","span":{"begin":2401,"end":2405},"obj":"CHEBI:22918"},{"id":"T3141","span":{"begin":2516,"end":2525},"obj":"GO:0008152"},{"id":"T3142","span":{"begin":2780,"end":2785},"obj":"UBERON:0002107"},{"id":"T3143","span":{"begin":2789,"end":2810},"obj":"GO:0006520"},{"id":"T3144","span":{"begin":2857,"end":2862},"obj":"NCBITaxon:9606"},{"id":"T3145","span":{"begin":2863,"end":2868},"obj":"UBERON:0002107"},{"id":"T3146","span":{"begin":2928,"end":2933},"obj":"UBERON:0002107"},{"id":"T3147","span":{"begin":2991,"end":2996},"obj":"UBERON:0002107"},{"id":"T3148","span":{"begin":3128,"end":3145},"obj":"CHEBI:35705"},{"id":"T3149","span":{"begin":3146,"end":3151},"obj":"CHEBI:23888"},{"id":"T3150","span":{"begin":3253,"end":3258},"obj":"UBERON:0002107"},{"id":"T3151","span":{"begin":3305,"end":3311},"obj":"UBERON:0002107"},{"id":"T3152","span":{"begin":3615,"end":3621},"obj":"NCBITaxon:33208"},{"id":"T3153","span":{"begin":3857,"end":3863},"obj":"NCBITaxon:33208"},{"id":"T3154","span":{"begin":3875,"end":3880},"obj":"NCBITaxon:9606"},{"id":"T3155","span":{"begin":3962,"end":3968},"obj":"NCBITaxon:33208"},{"id":"T3156","span":{"begin":4000,"end":4005},"obj":"NCBITaxon:9606"},{"id":"T3157","span":{"begin":4027,"end":4048},"obj":"CHEBI:23995"},{"id":"T3158","span":{"begin":4050,"end":4053},"obj":"CHEBI:23995"},{"id":"T3159","span":{"begin":4070,"end":4075},"obj":"NCBITaxon:10088"},{"id":"T3160","span":{"begin":4106,"end":4111},"obj":"NCBITaxon:9606"},{"id":"T3161","span":{"begin":4189,"end":4195},"obj":"GO:0008380"},{"id":"T3162","span":{"begin":4189,"end":4200},"obj":"SO:0000162"},{"id":"T3163","span":{"begin":4208,"end":4221},"obj":"GO:0005739"},{"id":"T3164","span":{"begin":4208,"end":4253},"obj":"PR:000004683"},{"id":"T3165","span":{"begin":4261,"end":4265},"obj":"SO:0000704"},{"id":"T3166","span":{"begin":4377,"end":4382},"obj":"NCBITaxon:10088"},{"id":"T3167","span":{"begin":4401,"end":4406},"obj":"NCBITaxon:9606"},{"id":"T3168","span":{"begin":4478,"end":4489},"obj":"SO:0000704"},{"id":"T3169","span":{"begin":4501,"end":4507},"obj":"NCBITaxon:39107"},{"id":"T3170","span":{"begin":4601,"end":4606},"obj":"NCBITaxon:9606"},{"id":"T3171","span":{"begin":4678,"end":4697},"obj":"PR:000006300"},{"id":"T3172","span":{"begin":4729,"end":4738},"obj":"UBERON:0000922"},{"id":"T3173","span":{"begin":4729,"end":4743},"obj":"_FRAGMENT"},{"id":"T3174","span":{"begin":4749,"end":4754},"obj":"CL:0002322"},{"id":"T3175","span":{"begin":4745,"end":4747},"obj":"_FRAGMENT"},{"id":"T3176","span":{"begin":4835,"end":4837},"obj":"PR:000006300"},{"id":"T3177","span":{"begin":4838,"end":4842},"obj":"SO:0000704"},{"id":"T3178","span":{"begin":4920,"end":4926},"obj":"NCBITaxon:39107"},{"id":"T3179","span":{"begin":5026,"end":5030},"obj":"SO:0000704"},{"id":"T3180","span":{"begin":5034,"end":5043},"obj":"CL:0000034"},{"id":"T3109","span":{"begin":292,"end":296},"obj":"CHEBI:22918"},{"id":"T3110","span":{"begin":674,"end":678},"obj":"CHEBI:22918"},{"id":"T3111","span":{"begin":682,"end":687},"obj":"UBERON:0000178"},{"id":"T3100","span":{"begin":11,"end":16},"obj":"NCBITaxon:4022"},{"id":"T3101","span":{"begin":23,"end":28},"obj":"UBERON:0001088"},{"id":"T3102","span":{"begin":49,"end":56},"obj":"SO:0000704"},{"id":"T3103","span":{"begin":107,"end":116},"obj":"CHEBI:25754"},{"id":"T3104","span":{"begin":142,"end":155},"obj":"GO:0005739"},{"id":"T3105","span":{"begin":168,"end":175},"obj":"GO:0032991"},{"id":"T3106","span":{"begin":196,"end":205},"obj":"MOP:0000568"},{"id":"T3107","span":{"begin":240,"end":250},"obj":"CHEBI:25754"},{"id":"T3108","span":{"begin":264,"end":290},"obj":"CHEBI:22918"},{"id":"T3112","span":{"begin":698,"end":709},"obj":"UBERON:0006314"},{"id":"T3113","span":{"begin":986,"end":990},"obj":"CHEBI:22918"},{"id":"T3114","span":{"begin":1120,"end":1125},"obj":"UBERON:0000178"},{"id":"T3115","span":{"begin":1126,"end":1130},"obj":"CHEBI:22918"},{"id":"T3116","span":{"begin":1357,"end":1370},"obj":"GO:0017086"},{"id":"T3117","span":{"begin":1464,"end":1477},"obj":"CHEBI:50488"},{"id":"T3118","span":{"begin":1492,"end":1494},"obj":"PR:000006300"},{"id":"T3119","span":{"begin":1503,"end":1516},"obj":"CHEBI:50488"},{"id":"T3121","span":{"begin":1590,"end":1591},"obj":"_FRAGMENT"},{"id":"T3122","span":{"begin":1602,"end":1610},"obj":"PR:000004686"}],"relations":[{"id":"R339","pred":"_lexicallyChainedTo","subj":"T3122","obj":"T3121"},{"id":"R340","pred":"_lexicallyChainedTo","subj":"T3128","obj":"T3127"},{"id":"R341","pred":"_lexicallyChainedTo","subj":"T3174","obj":"T3173"},{"id":"R342","pred":"_lexicallyChainedTo","subj":"T3174","obj":"T3175"}],"text":"Background\nMaple Syrup Urine Disease (MSUD) is a genetic disorder caused by a deficiency of branched-chain keto acid dehydrogenase (BCKDH), a mitochondrial multienzyme complex responsible for the oxidative decarboxylation of branched-chain keto acids derived from branched-chain amino acids (BCAA), leucine, isoleucine and valine (for review, see: [1]).\nPatients with MSUD, depending on the mutation, show variable degrees of enzyme deficiency leading to several different clinical phenotypes [1]. Approximately 75% of MSUD patients have the classic form of the disease with BCKDH activity in the range of 0–2% of normal [1]. These patients show markedly elevated levels of BCAA in blood and other body fluids [1]. Besides the classic form, there are other variants of the disease. Patients with the intermediate form of the disease show BCKDH activity in the range of 3–30% of normal. In such patients the onset of the disease is delayed, but there are persistently elevated levels of BCAA [1]. Patients with the intermittent form of MSUD show BCKDH activity in the range of 5–20% and during the asymptomatic phase the blood BCAA levels are normal [1]. The overall incidence of MSUD in the general population is 1:185,000 [1], and in certain population groups, such as Mennonites of Pennsylvania, the incidence is estimated to be as high as 1:176 [2].\nThe BCKDH complex, the deficient enzyme in MSUD, consists of three catalytic proteins, a decarboxylase (E1), a dihydrolipoyl transacylase (E2), and a dihydrolipoyl dehydrogenase (E3). The E1 component is a heterotetramer composed of two α and two β subunits [3]. The E1 and E2 components are specific to BCKDH, whereas E3 is also used by pyruvate and α-ketoglutarate dehydrogenase complexes [3] and the glycine cleavage system [4]. BCKDH is also associated with two regulatory proteins, a specific kinase and a phosphatase which regulate the activity of this enzyme through a phosphorylation (inactivation) and dephosphorylation (activation) cycle of the E1 α subunit [5,6]. Mutations in the genes of the E1 and E2 subunits of BCKDH have been described, however, the majority of MSUD mutations identified thus far are in the E2 subunit [1,7]. To date, cases of MSUD have not been associated with defects in the regulatory kinase and phosphatase [1].\nCurrent management of MSUD patients relies on a strict lifelong dietary restriction of protein or BCAA [1,8]. Such a dietary management of the disease, however, is not entirely satisfactory especially in times of metabolic decompensation due to infection, injuries and other stressors. In spite of dietary intervention, there is significant mortality associated with MSUD and there is a high incidence of mental retardation in survivors [9].\nBecause of the central role of the liver in amino acid metabolism and moderate/high levels of BCKDH activity in human liver [10-12], a few cases of MSUD have recently been treated by liver transplantation [13-16]. While the short-term outcome of liver transplantation is encouraging, long-term effects of this approach are not known. However, these patients are now required to take immunosuppressant drugs for the rest of their lives, often with undesirable side effects. Moreover, the cost associated with liver transplantation and the availability of donor livers are additional limiting factors for the practicality of treatment of this disease.\nBecause of the current unsatisfactory options for the treatment of MSUD, there is a need for improved therapies to combat this disease. An obstacle to developing novel treatments for MSUD has been the lack of a suitable animal model to perform necessary preclinical studies. Although a Hereford calf model with MSUD has been described [17-19], this model is neither readily available nor practical to perform preclinical studies. Furthermore, comparison of this animal model with human MSUD has shown some differences in the pathology of the disease [1], making this animal a less desirable model for the human disease.\nRecently, a N-ethyl-N-nitrosourea (ENU)-induced mutant mouse that phenotypically resembles human MSUD has been described [20]. However, the mutation in this model disrupts a splice site in the mitochondrial branched-chain aminotransferase (BCAT) gene, not in BCKDH, the deficient enzyme in MSUD. Because the mutation is not in BCKDH, the validity of this mutant mouse line for modeling human MSUD is questionable.\nThe objective of the present study was to create genetically engineered murine models of MSUD that mimic the pathology of the classic and intermediate variant forms of the human disease. The classic model was created by targeted inactivation of the E2 subunit of BCKDH by homologous recombination in embryonic stem (ES) cells. The model of intermediate MSUD was created by partial transgenic rescue of the E2 gene knockout. This report describes the generation and characterization of these murine models of MSUD. These models will allow for the development of novel treatment approaches, such as gene or stem cell therapies, to ultimately cure MSUD."}