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Anaplerosis

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Metabolic Map Detailed

Core Pathways
TCA Cycle
8 metabolites · Citrate → Oxaloacetate
Rate-limiting: Isocitrate DH
Glycolysis
Glucose → Pyruvate → Acetyl-CoA
Rate-limiting: PFK-1
Electron Transport Chain
CI → CoQ → CIII → CytC → CIV → ATP Synthase
NADH/FADH₂ → O₂ → H₂O + ATP
β-Oxidation
Fatty acids → Acetyl-CoA (spiral)
VLCAD / LCHAD / MCAD
Flux Pathways
Anaplerotic Input ↗
Feeds carbons INTO the TCA cycle
PC, GDH, Triheptanoin
Cataplerotic Exit ↘
Drains intermediates for biosynthesis
PEPCK, ACLY, ALAS, CAD
Amino Acid Input
Protein catabolism → TCA intermediates
GDH, GOT, BCKDH, ALT
Interventions
Cofactors / Supplements
CoQ10, B₂, Carnitine, B₁, B₇, ALA
Dashed lines → targets
Symbols
🔴 Disease link
⚡ Energy output
★ FDA-approved
⬡ Ball-and-stick model
● Red glow = blocked
● Blue = depleted

⚗ Intervention Layer

Click a TCA enzyme in block mode to simulate Le Chatelier effects
⚠ ENZYME BLOCK MODE — Click a TCA metabolite to simulate deficiency
Understanding

What is Anaplerosis?

From Greek ana (up) + plerotikos (to fill). Anaplerosis is the replenishment of TCA cycle intermediates — the metabolic reactions that keep the engine running when its parts are consumed for other purposes.

🔄

The Problem

TCA cycle intermediates are constantly being pulled away — for gluconeogenesis, amino acid synthesis, heme production, and fatty acid building. Without replenishment, the cycle stalls and energy production collapses.

The Solution

Anaplerotic reactions feed new carbon skeletons back into the cycle. Pyruvate carboxylase (→ OAA), glutaminolysis (→ α-KG), and odd-chain fatty acid oxidation (→ succinyl-CoA) are the three major routes.

💊

The Medicine

Triheptanoin (Dojolvi®) is the first and only FDA-approved anaplerotic therapy. Its C7 fatty acid chains produce propionyl-CoA, which becomes succinyl-CoA — directly replenishing the cycle where it matters most.

🧬

When It Breaks

TCA cycle enzyme deficiencies cause devastating neurological disease. But heterozygous loss of the same genes — SDH, FH, IDH — drives cancer through oncometabolite accumulation. The cycle connects rare disease to oncology.

🫁

Every Breath

Two carbons enter each turn as acetyl-CoA. Two leave as CO₂ — becoming the air you exhale. The TCA cycle is why you breathe. It has been spinning in your cells since the moment you were conceived.

🌍

Ancient

The TCA cycle predates oxygen in Earth's atmosphere. It likely ran in reverse in early anaerobic life, fixing CO₂ into organic molecules. It is one of the oldest metabolic pathways — perhaps 4 billion years old.

The Cycle

Eight Steps, One Turn

Each turn of the TCA cycle extracts the energy stored in acetyl-CoA's carbon bonds, capturing it as electron carriers (NADH, FADH₂) and direct energy (GTP), while releasing CO₂.

1

Oxaloacetate + Acetyl-CoA → Citrate

Citrate Synthase (CS)

The gateway reaction. A 4-carbon molecule grabs a 2-carbon fuel to form 6-carbon citrate.

Condensation
2

Citrate → Isocitrate

Aconitase (ACO2)

A molecular rearrangement via dehydration-rehydration. Aconitase's iron-sulfur cluster is sensitive to oxidative stress.

ACO2 deficiency
3

Isocitrate → α-Ketoglutarate

Isocitrate Dehydrogenase (IDH3)

First carbon exhaled as CO₂. First NADH captured. IDH mutations produce oncometabolite D-2-HG.

NADHCO₂IDH cancers
4

α-Ketoglutarate → Succinyl-CoA

α-KG Dehydrogenase Complex (OGDH/DLD)

Second CO₂ released. Another NADH. DLD is shared with PDH — one mutation disables both.

NADHCO₂DLD deficiency
5

Succinyl-CoA → Succinate

Succinyl-CoA Synthetase (SUCLA2/SUCLG1)

GTP born through substrate-level phosphorylation. This is where triheptanoin delivers its payload.

GTPmtDNA depletion
6

Succinate → Fumarate

Succinate Dehydrogenase / Complex II (SDHA/B/C/D)

The only enzyme in BOTH TCA and electron transport. SDH IS Complex II.

FADH₂SDH tumors
7

Fumarate → Malate

Fumarase (FH)

Biallelic FH loss = devastating IEM. Heterozygous = HLRCC with aggressive renal cancer.

FH deficiency / HLRCC
8

Malate → Oxaloacetate

Malate Dehydrogenase (MDH2)

The wheel completes. Final NADH captured. OAA is regenerated, ready for another turn.

NADHMDH2 deficiency

Net Yield Per Turn

3
NADH → ~7.5 ATP
1
FADH₂ → ~1.5 ATP
1
GTP → 1 ATP
2
CO₂ → exhaled

~10 ATP per turn · 2 turns per glucose · ~30–32 ATP total from oxidative phosphorylation

Treatment

Anaplerotic Therapies

Therapies that replenish TCA cycle intermediates — from the only FDA-approved anaplerotic drug to investigational compounds.

L-Glutamine (Endari®)

FDA-approved for sickle cell disease

Glutamine → glutamate → α-ketoglutarate via GDH. Directly replenishes TCA. Dominant anaplerotic pathway in rapidly dividing cells and cancer.

→ α-Ketoglutarate

Odd-Chain Fatty Acids

C15:0 (pentadecanoic acid) · C17:0 (heptadecanoic acid)

Found in dairy fat and ruminant meat. β-oxidation yields propionyl-CoA → succinyl-CoA. Same mechanism as triheptanoin.

→ Succinyl-CoA

α-Ketoglutarate (AKG)

Ornithine-AKG · Calcium-AKG

Directly enters TCA as α-ketoglutarate. Used in burn patients and surgical recovery. Emerging interest in aging research.

→ α-Ketoglutarate

Branched-Chain FAs

dMC7 · AdMC7 · dMC9 · Investigational

Medium branched-chain fatty acids. Preclinical data shows improved TCA intermediate profiles vs. heptanoate in VLCAD/LCHAD/TFP/CPT-II fibroblasts.

→ Succinyl-CoA

Pyruvate Carboxylase

Endogenous enzyme · Biotin-dependent

The PRIMARY anaplerotic reaction: pyruvate + CO₂ + ATP → oxaloacetate. Allosterically activated by acetyl-CoA. When deficient, the entire TCA cycle starves.

→ Oxaloacetate
Clinical Genetics

When the Cycle Breaks

TCA cycle enzyme defects cause some of the rarest and most severe inherited metabolic diseases. Partial loss of the same genes drives cancer — connecting rare disease to oncology.

SDH Tumor Syndromes

SDHA · SDHB · SDHC · SDHD · SDHAF2

Succinate accumulation inhibits α-KG-dependent dioxygenases, causing DNA hypermethylation and pseudo-hypoxia. Drives paraganglioma, pheochromocytoma, GIST, and renal cell carcinoma.

TCA Step: Succinate → Fumarate

IDH-Mutant Cancers

IDH1 (cytoplasmic) · IDH2 (mitochondrial)

Neomorphic mutations convert α-KG to D-2-hydroxyglutarate — an oncometabolite rewiring epigenetics. Found in >70% of low-grade gliomas, ~20% of AML. FDA drugs: ivosidenib, enasidenib, vorasidenib.

TCA Step: Isocitrate → α-Ketoglutarate

Fumarase Deficiency / HLRCC

FH · OMIM 606812

Biallelic: severe encephalopathy, fumaric aciduria. Heterozygous: hereditary leiomyomatosis + aggressive type 2 papillary RCC. Fumarate = oncometabolite.

TCA Step: Fumarate → Malate

Succinyl-CoA Ligase Deficiency

SUCLA2 · SUCLG1 · OMIM 612073 / 245400

SUCLA2: encephalomyopathy with mtDNA depletion and methylmalonic aciduria. SUCLG1: fatal infantile lactic acidosis. The link between the TCA cycle and the mitochondrial genome.

TCA Step: Succinyl-CoA → Succinate

Pyruvate Carboxylase Deficiency

PC · OMIM 266150

Three types: Type A (infantile), Type B (neonatal, severe), Type C (benign). When the primary anaplerotic enzyme fails, OAA cannot be replenished, gluconeogenesis halts.

Anaplerotic Input: Pyruvate → Oxaloacetate

LC-FAOD Spectrum

ACADVL · HADHA · HADHB · CPT1A · CPT2 · SLC25A20

VLCAD, LCHAD, TFP, CPT-I/II, CACT. Cannot generate acetyl-CoA from long-chain fats. TCA intermediates deplete. Triheptanoin (Dojolvi) is FDA-approved for these conditions.

TCA Impact: Acetyl-CoA + anaplerotic depletion
Therapeutic Biochemistry

The Mitochondrial Cocktail

When oxidative phosphorylation fails, targeted cofactors can bypass bottlenecks, shuttle electrons around damaged complexes, and scavenge the reactive oxygen species that destroy from within. This is anaplerotic medicine.

Le Chatelier in the Mitochondria

When an enzyme is blocked — by genetic deficiency or pharmacologic inhibition — substrates accumulate upstream and products deplete downstream. This is Le Chatelier's principle applied to living biochemistry. The TCA cycle doesn't just slow: it distorts, creating metabolite imbalances that drive disease. Succinate accumulation from SDH loss activates pseudo-hypoxia. Fumarate accumulation from FH loss rewrites the epigenome. Understanding these shifts is the foundation of metabolic therapeutics.

Toggle "Enzyme block mode" in the 3D viewer above, then click any TCA metabolite to see the cascade.