CU Biochem Course and NAMA Flashcards
What is the difference between classic PKU, mild PKU< and hyperpheylalaninemia?
Untreated Phe levels:
>1200umol/L - classic PKU
600-1200umol/L - mild PKU
<600umol/L - hyperphe
Mousy odored urine, light hair, eczema, developmental delay, white matter hyperintensities
Untreated PKU
- treatment needs to start within 1 month of life
- “pseudoleukodystrophy”
What is pegvaliase
Enzyme substitution for PAH deficiency (PKU)
- plant enzyme (phe ammonia lyase)
- Goal to enhance Phe tolerance and normalize diet
- still need tyrosine
- Approved for 16+
Elements of PKU diet
Low Phe
Supplement tyrosine and tryptophan (share same large neural AA transporter as Phe)
What is a goal of sapropterin trial in PKU?
20mg/kg/day
Infant: 24 hour trial -> >30% reduction in Phe = positive (no dietary changes)
Adult: 48hr to 30 day trial
Mutatons in which subunit of BCKDH can be thiamine (B1) responsive?
E2 subunit
What is the role of IV destroxe (glucose) in metabolic decompensation?
- Prevent gluconeogenesis that drives protein breakdown
- Provide calories
- Drives insulin production (anabolic)
Tyrosine aminotransferase deficiency
Tyrosinemia type 2
PAA: high Tyr, Phe
UOA: 4-OH phenylpyruvate/lactate/acetate
- corneal lesions, hyperkeratosis
UOA: 4-OH phenylpyruvate, 4OH phentyllactate, 4OHphentylacetate
Tyrosinemia type II
Tyrosine aminotransfearse deficiency
TAT gene
PAA: high Tyr, Phe
4-hydroxyphentylpyruvic dioxygenase deficiency
Tyrosinemia type III
HPD gene
- PAA: high Tyr, Phe
- Uncertain clinical significance, may have keratitis
Renal fanconi, liver/renal disease, porphyric crisis, hepatocellular carcinoma
Tyrosinemia type I
Fumarylacetoacetate hydrolase deficiency
FAH
- High AFP, SUAC, Met (liver disease)
high AFP, methionine, aaminolaevulinic acid (ALA)
Tyrosinemia type I
FAH
What are the 3 presentations for tyrosinemia type I?
- Hepato: early severe liver disease
- Renal: rickets (renal fanconi)
- Acute intermittent: porphyria like attacks
What are the toxic compounds in tyrosinemia type I?
FAH deficiency
- Fumarylacetoacetate, Maleylacetoacetate, Succinylacetone
Which steps in porphyrin synthesis does SUAC inhibit?
ALAD and HMBS
What is 2-nitro-4-trifluoromethylbenzoyl - 1,3- cyclohexane-dione used to treat?
Tyrosinemia type I
2-nitro-4-trifluoromethylbenzoyl - 1,3- cyclohexane-dione
NTBC = nitsinone
- inhibits 4-OHphentylpyrvic acid dioxygenase
- Goal: blood level >40 (enzyme inhibition 99%); SUAC suppression
- restrict Phe/Tyr to goal <600 umol/L in blood
corneal crystals, plantarpalmar hyperkeratosis with pits
Type II or III tyrosinemia
TAT (tyrosine aminotransferase), HPD (hydroxyphenylpyruvate dioxygenase)
- May also have ID/seizures if untreated
If NBS shows high Tyr and normal SUAC, what should you do next?
Repeat at 6 weeks -> if normal this is transient tyrosinemia of newborn
-> if abnormal work up tyrosinemia type II and III
What vitamin will 50% of Cystathione beta syntase deficiency respond to?
B6 - pyridoxine
What is treatment of homocystinuria is B6 unreponsive?
Folate (B9)
B12 if deficient
Low protein (met)
Betaine (alternate remethylation pathway)
Which UCD has fair fragility (Trichorrhexis nodosa) + rash?
ASL deficiency
- also increased liver disease
Which UCD enzymes are in the mitochondria?
N-acetylglutamate synthetase
Carbamoyl phsophate synthetase
Carbonic anhydrase 5A (CAVA) - Makes bicarb necessary for carboxylases (including CPS)
Ornithine transcarbamylase
Why does citrulline bruild up in citrin deficiency?
Aspartate is stuck and mitochondria and unable to conjugate with citrulline in cytoplasm (ASS) to form arginosuccinate.
Why is Lys high in OTC deficiency?
Shortage of 2-oxoglutarate
Gold standard testing for OTC in females
Liver enzyme funciton testing
- Molecular testing cannot find mutation in 10%
- Orotic acid/glutamine may be normal
Why does Carbonic anhydrase 5-A deficiency cause hyperammonemia?
CAVA synthesizes bicarb in mitochondria used by CPSI -> Proximal UCD
What is phenylbutyrate?
Prodrug to phentylacetate -> conjugates to glutamine
Which UCD presents with spastic diplegia and intellectual disability?
Arginase deficiency
+ growth arrest
- check PAA when working up DD!
What are the cofactors for glycine cleavage system?
Lipoate and pyridoxal-Phosphate
- Deficiency can cause variant NKH
high CSF/serum glycine, high homocysteine, high MMA
HCFC1 - Cobalamin X
- XL
- Transcription factor for glycine cleavage system and Cbl pathway
What drugs should be avoided in NKH?
Valproic acid and vigabatrin -> inhibit residual enzyme activity
(both can cause iatrogenic glycine elevation)
98% of NKH are caused by variants in what 2 genes?
GLDC and AMT
P and T protein
What is treatment for NKH?
Benzoate (conjugates with glycine to form hippurate) -> goal plasma glycine 120-300uM
Glycine restriction
Ketogenic diet (glycine is gluconeogenic substrate)
NMDA blockade: Dextromethorphan, ketamine
What is serine synthesized from?
3-phosphoglycerate (glycolysis intermediate)
3PG Dehydrogenase -> Phosphserine aminotransferase -> phosphosering phosphatase
CSF: low glycine, low serine
Serine synthesis defect
PGDH, PSAT, PSPH
- Severe hypomyelination and epilepsy
What is the treatment for neu laxova syndrome?
Serine supplementation + glycine supplementation
What is the vulnerable window in GA1
3 months - 2 years
- Up to 6yo can be affected
High Glutaric acid, 3-OH glutaric acid
GA1
GCDH deficiency
What is C5DC?
Glutaryl carnitine
- look for GA1
What is treatment for GA1?
1) avoid catabolism
2) low lysine/trytophan
3) Carnitine
What does 2-aminoadipic semialdehyde dehydrogenase deficiency cause?
Pyridoxine dependent epilepsy
ALD7A1 - antiquitin
High aminoadipid acid semialdehyde, pipecolic acid, 6-oxo-pipecolate
pyridoxine dependent epilepsy
ALD7A1
What is the second most common B6 responsive epilepsy? (after ALDH7A1)
Pyridoxine 5 phosphate oxidase deficiency
PNPO
- Converts pyridoxine to active form
- Normal cognitive development (not true in ALDH7A1)
What is the treatment for alpha-aminoadipic semialdehyde DH deficiency or pyridoxamine-5-phosphate oxidase deficiency?
Pyrixodine depedent epilepies (ALDH7A1 and PNPO)
- Will always respond to pyridoxal phosphate (active form)
- might repsond to pyridoxine
Which pathways are mostly affected in riboflavin deficiency?
ETFDH (GA2)
FAOD
BCAA metabolism
Complex I
Axonal neuropathy, hearing loss, bulbar weakness, myopathy
Riboflavin transporter defects
RFVT2 and RFVT3
Brown-Vialetto- Van Laere
- Tx: Riboflavin
Low 5-methyltetrahydrofolate in CSF
Folate transporter deficiency (FOLR1) or Kearns-Sayre
- Ataxia, epilepsy, demyelination
megaloblastic anemia, diabetes, SNHL in infancy
Thiamin-responsive megaloblastic anemia
SLC19A2
3 thiamine reponsive encephalopathies
TPK1
SLC19A3
SLC25A19 (also severe congenital microcephaly)
What does G6Pase translocase deficiency cause?
GSD1b
- GSD1 + neurtropenia
GSD1a is caused by G6Pase (most common GSD)
Are glucagon levels low or high in GSD1?
Elevated
Glucagon drives gluconeogenesis and glycogenolysis, both are defective in G6Pase deficiency
- Insulin is low until fed
Doll like face, buttock fat pad accumulation, hepatomegaly, nephromegaly, hypotonia, bleeding tendency
GSD1
- No nephro/hepatomegaly in CDG-PMM2
Compare Von Gierke and Cori disease
GSD1 (G6Pase) vs GSD3 (amylo-1,6-glucosidase)
- GSD3 has milder hypoglycemia (gluconeogenesis intact), but more early liver dysfunction
- Give high protein diet for GSD3 (glucogeogenesis)
What is the mechanism of injury in Anderson disease?
GSD4 - a-1,4-glucan-6-glucosyltransferase deficiency (branching)
- Unbanched glycogen (polyglucosan) build -> toxic -> hepatosplenomegaly
- other GSDs have branched glycogen buildly
Liver pathology findings for Andersen’s disease
GSD4 - branching enzyme
- Unbranched glycogen (polyglusocan) accumulation
- path: PAS stain positive, diastase-resistant with amylopectin-like inclusions
Which GSDs have postprandial hyperglycemia?
GSD0 - unable to store BG as glycogen
What can patients with fanconi Bickel syndrome get that GSD1 patients canno?
Fructose
- Fanconi bickel = glucose/galactose transporter defect
- in GSD1 fructose cannot be used because it is convered into G6P
Treatment for tauri disease?
Avoid sugars/glucose - no NOT give D10 when sick
- Glucose is high but not sensed by mitochondrial/cannot be used for glycolysis.
- FAO inhibited by high glucose levels
In which GSDs should you avoid giving excess glucose?
GSD4 (Andersen) and GSD9 (Tauri)
- Andersen -> more glucose -> more polyglucosan; give proteins to promote gluconeogensis instead
- Tauri -> Glycolysis is blocks, reduce glucose to release inhibition on FAO
microcephaly, thin upper lip, long philtrum, wide nasal bridge, DD, leigh syndrome
Pyruvate dehydrogenase deficiency
Most common X linked
- Fetal alcohol syndrome-like face + leigh syndrome
- high pyruvate -> high lactate + alanine
- Lacate/pyruvate ratio low
Treatment for PDH deficiency
Ketogenic diet, thiamine, carnitine, lipoic acid
Name the 3 glyconeogenic enzymes
Pyruvate Carboxylase, Fructose 1,6 bis phosphatase, Glucose-6-phosphatase
Cystic brain lesions, FTT, metabolic acidosis
PAA: High Cit, Ala, Lys, Pro
UOA: 2-oxoglutaric acid (aKG)
High lacate/pyruvate with increased ratio >20
high acetoacetate/betahydroxy-butyrate ratio
High ammonia
Pyruvate Carboxylase deficiency
- Biotin Dependent
- TCA cycle and gluconeogenesis affected
High lacate -> high ala, proline (proline oxidase suppressed)
Low OAA (precursor) -> low aspartate and glutamate
Low aspartate -> high citrulline and ammonia
Compare F1,6BP deficiency with HFI
- F1,6BP can tolerate more fructose
- F1,6BP presents more like GSD1 - F1,6BP buildup inhibits G6Pase
- FBP: Pseudohypertriglyceridemia, high glycerol, glycerol 3 phosphate
What is the pathophys of HFI?
Aldolase B deficiency
Toxic accumulation of F1P -> impaired gluconeogensis and glycogenolysis -> lactic acidosis, hypolgycemia, liver failure
Promoter deletion + N314D/Q188R
Biochemical (Duarte) Galactosemia
GALT (~25% activity)
UOA: high glycerol-3-phosphate, glycerol
high TG
high alanine, ketones
Fructose 1,6 BP
Glycerol-3-phosphate is not seen in glycerolkinase
Contrast HFI and FBPase deficiency
HFI: childrne have no cavities (Aversion to sweets), FTT with fructose ingestion, RTA
FBPase: can tolerate fructose, no renal involvement, UOA with glycerol/glycerol-3-phosphate
Q188R
K285N
L195P
classic galactosemia
GALT (<1% activity)
- early e.coli sepsis, FTT, liver disease
+ Premature ovarian insufficiency and DD
S125L
Variant galactosemia
GALT (1-10% activity)
Same early presentation (e-coli sepsis, FTT, liver disease etc)
- Normal development/reproduction
Urine polyol analysis: high ribitol, D-arabitol, erythritol, sedoheptulose
Transaldolase
TALDO
Liver, HSM, anemia, thrombocytopenia, congenial anomalies
Urine polyol: high ribitol, D-arabitol
Ribose-5-P Isomerase
RPIA
- Leuckoencephalopathy, ataxia, neuropathy
What are the steps of ß-Oxidation?
1) Dehydrogenase (create 2,3 double bond)
2) hydratase (add 3-OH)
3) dehydroganse (create 3 double bonded O)
4) thiolase (remove ketone)
Treatment for FAODs
- low fat diet
- avoid catabolism
- triheptanoin (Long chain and above) - can freely diffuse into mitochondrial and bypass carnitine shuttle/long chain breakdown
- Carnitine (allows urine clearance of FA)
What is OCTN2 deficiency?
Primary carnitine deficiency (Carnitine transporter)
(SLC22A5)
- carnitine cannot be reabsorbed in kidney -> unavailable in liver/muscle/heart
- Presents with hepatic encephalopathy, cardiac, hypoglycemia, rhabdo
What should be done if a baby is NBS positiove for low carnitine but has normal plasma F/T carnitine testing?
Screen for mother with primary carnitine deficiency
- Carnitine comes from breastmilk for young infants
- Mothers w/ carnitine deficiency will have NBS positive child and are at risk for sudden death
UOA: Hexanoylglycine
MCAD
ACADM
ACP: high C6, C8, C10:1
Why are dicarboxylic acids elevated in urine in FAOD?
Long chain FAs build up in cytoplasm go into microsomes -> gets converted into dicarboxylic acids
What is the next step if NBS is positive with elevated C14:1 but plasc ACP is normal?
VLCAD gene sequencing (ACP may be normal between episodes)
FAOD + neuropathy, RP, and maternal preeclampia/HELLP/fatty liver
LCHAD/TFP
- hydroxylated FAs can flow into mom and accumulate
- TFP also modified cardiolipin (may explain extra symptoms)
if NBS is high for C16OH and ACP/UOA are normal, what is the next step?
Sequencing for LCHAD/TFP - cannot rule out wth ACP alone
UOA: High ethylmalonic acid, n-butrylglycine, and methylsuccinic acid
- SCAD
- Isobutyryl glycinuria (+ high isobutyrylgylcine)
- Ethylmaloni encephalopathy (+ persistent lactic acidosis) - ETHE
What is the final electron acceptor of dehydrogenases?
ETFDH
- Causes GA2 -> mulitiple DH issues -> multiple chain length elevations of ACP
- Riboflavin is cofactor
What are the main ketogenic AAs?
Leucine, isoleucine, lysine
Hypoketotic hypoglycemia, dicarboxylic aciduria, 4-hydroxy-6-methyl-2-pyrone
- normal ACP
3-Hydroxy-3-emethly-glutaryl-CoA synthase deficiency (HMG-coa Synthase)
UOA: 3-hydroxy-3-methylglutaric acid, 3-methylglutaric acid
- hyperammonia, hypoketotic hypoglycemia
2-Hydroxy-3-methlyglutayl-CoA Lyase deficiency (HMG-coa Lyase)
- high HMG and MG (3-hydroxy-3-methylglutaric acid, 3-methylglutaric acid)
- C6DC (3 methylglutaryl-carnitine), C5OH (3-OH-isovaleryl carnitine)
high D-3-hyroxybutyrate, tiglylglycine, 2-methyl-3-hydroxybutryrate, 2-methylacetoacetate
beta-ketothiolase (T2)
- ACAT1
- High ketones during illness + isoleucine intermediates
High ketones with normal dicarboxylic acids
Disorders of ketolysis
- physiological ketosis comes from FAO -> thus dicarboxylic acids (adipid, suberic, and sebacic) also build up
- HIgh ketones with normal dicarboxylic acids -> usually SCOT deficiency
High ketones during fed and fasting state
Succinyl-Coa - oxoacid-Coa transferase (SCOT)
SCOT/OXCT1 gene
- Do not see elevated dicarboxylic acids (adipic, suberic, sebacic) which are seen in physiological ketosis
DDx: Monocaroboxylic transporter 1 (MCT1) deficinecy -> ketone transporter
What is Monocarboxylic Transporter 1 (MCT1) deficiency?
Ketone transporder defect
SLC16A1
- looks similar to SCOT -> episodic ketoacidosis
- Increased acetoacetate + 3 OH butyric acid, normal dicarboxylic acids
Renal fanconi, renal failure, cornea crystals, hypothyroidism, hypogonadism, myopathy
Cystinosis
Lysosomal Cystine transporter
- Tx Cysteamine eyedrops + Tablets
- Cysteine (Cystine + Cysteamine) has own transporter; another cysteine can be added and use Lysine transporder
GLA N215S
Cardiac Variant Fabry
GLA A143T
VUS Fabry
- Most common on NBS
Alkylglycerone 3-phosphate acyltransfearse deficiency
GNPAT - RCDP
Alkylglycerone 3-phosphate synthase deficiency
AGPS
- RCDP
Most common cause of RCDP
PEX7
- Low plasmalogen, also high phytanic acid
Fatty Alcholol Reductase deficiency
RCDP without RCDP
PEX5L deficiency
RCDP
- mutation in specific long isoform of PEX5 - mutations in exon 9
- Results in abnormal PEX7 activity
Metaphyseal flaring, epiphyseal stippling, coronal vertebral clefts, mineralization of intervertebral disks
RCDP
- ARSE - XL -Male
- Sterol-8-isomerase - XL - Females (male lethal)
- Wafarin/VitK, EtOH, Phenytoin
high oxalate, glyoxalate
Primary hyperoxaluria type I
AGT
Alanine-glyoxylate aminotransferase - peroxisomal enzymes
- Renal stones + nephrocalcinosis
- Tx: Pyrixosone, Hydration, Liver/Kidney transplant
Neuropathy, loss of night vision, deafness, conduction defects, short metacarpals/metatarsals
Adult refsum disease
- Phytanol-CoA hydroxylase deficiency
- High phytanic (found in animal fats and plants), low pristanic acid
Which amino acides do glycosylation occur on?
N-linked - Asparagine
O-linked - Serine or threonine
Which organelles does glycosylation occur in?
Early - cytosol and ER (Type I isoelectric focusing pattern)
Late - Golgi (Type II pattern)
What methods to defect CDG is more sensitive and specific than isoelectric focusing?
Mass Spec
- Can see both N and O linked CDGs
- Can use flow-intection electrospray ionization to digest N-glycan off protein and get more specific results
What does phosphomannomutase do?
PMM2 -> converst mannose 6P to mannose 1P
Causes CDG
Which CDG causes protein losing enteropathy and hyperinsulinism with no neurological diasease
MPI - CDG1b
- treat w/ mannose
Which CDG is associated with elevated alk phos?
GPI anchor disorders
PIGA, PIGN, PIGO
- Seizures and DD
m.8993 T>G
Leigh syndrome - ATP6
m.3243 A>G
MELAS
rRNA leu
- Diabetes + hearing loss most common
- Complex I, III, IV deficiency
- Treat w/ Arg/Cit/Tauring
m.8344A>G
MERRF
tRNA Lys
+ lipomas
m.1555G>A
Hearing loss when exposed to aminoglycosides
(Mitocondria are derived from bacteria symbiotes!)
Homoplasmic m.11778 G>A (MT-ND4)
LHON
- Males > females
- incomplete penetrance
names 2 mtDNA deletion syndromes
Kearns-Sayre = CPEO + conduction block, RP, diabetes, renal, ID, hearing loss
Pearson Syndrome = KSS + sideroblastic anemia and exocrine pancreas dysfunction
SDH deficiency
AD paraganglioma + pheo
AR Leigh syndrome
Leigh syndrome + hypertrichosis/hirsutism
SURF1
- nuclear gene involved in complex IV assembly
- most common leigh gene
X-linked leigh syndrome
Pyruvate dehydrogenase deficiency
Tx - thiamine and ketogenic diet
- Males = mild mutations (severe mutations lethal)-> leigh syndrome, normal brain structure
- Females = het for severe mutations -> structural abnormalities, normal lactate (X inactivation)
POLG related disease
Alpers syndrome
- Multisystemic with many neurologicalm manifestations
- Polymerase Gamma
- VPA use can precipitate liver failure
- mtDNA depletion + Complesed I/III/IV deficiency
Steroid resistant nephrotic syndrome
CoQ10 synthesis defect
- ADCK4, PDSS2, COQ2, COQ6
Non-compaction cardiomyopathy, neutropenia, myopathy + high 3-methylglutaconic acid
Barth syndrome
TAZ (X Linked)
What does CblJ and CblF deficiency cause?
Combined HCy and MMA
- Cbl transport out of lysosome into cytosol where CblC is
- Serum B12 levels low - but NOT B12 deficiency
What does Methylmalonyl CoA Epimerase deficiency cause?
MMA and PA
- Slight elevation in MMA (only D-Methylmalonyl=CoA high) and proximal buildup of PA + 2-methylcitrate (secondary PA metabolite)
What should be ruled out if baby presents with NBS high MMA but no mutation is found?
Maternal B12 deficiency
What are the first steps after diagnosing sick child w/ MMA/PA?
- Determine B12 resonsiveness with hydroxocolbalamin
- Can try biotin if PA suspected (PA Carboxylase)
Bulls eye maculopathy
CblC (rarely CblD)
Bulls eye maculopathy can progress to RP and optic nerve atrophy despite treatment
Macocytic anemia, pancytopenia, high MMA, high HCy
Transcobalamin 2 deficiency
TCN2
- Treat with OH-Colbalamin injections
- Looks like neonatal leukemia/SCID
- TC2 colbalmin receptor deficiency tends to be milder/transient
What Cbl defects can have low B12 levels?
Cbl J and Cbl F
- abnormal lysosomal efflux
What is CblX?
HCFC1 and epi-cblC mutations
HCFC1 is X-linked
- Promoters/regulators of CblC expression