L-Leucine

Blood. 2012 Sep 13;120(11):2214-24.

L-Leucine improves the anemia and developmental defects associated with Diamond-Blackfan anemia and del(5q) MDS by activating the mTOR pathway.[Pubmed: 22734070 ]

Haploinsufficiency of ribosomal proteins (RPs) has been proposed to be the common basis for the anemia observed in Diamond-Blackfan anemia (DBA) and myelodysplastic syndrome with loss of chromosome 5q [del(5q) MDS]. We have modeled DBA and del(5q) MDS in zebrafish using antisense morpholinos to rps19 and rps14, respectively, and have demonstrated that, as in humans, haploinsufficient levels of these proteins lead to a profound anemia.
METHODS AND RESULTS:
To address the hypothesis that RP loss results in impaired mRNA translation, we treated Rps19 and Rps14-deficient embryos with the amino acid L-Leucine, a known activator of mRNA translation. This resulted in a striking improvement of the anemia associated with RP loss. We confirmed our findings in primary human CD34⁺ cells, after shRNA knockdown of RPS19 and RPS14. Furthermore, we showed that loss of Rps19 or Rps14 activates the mTOR pathway, and this is accentuated by L-Leucine in both Rps19 and Rps14 morphants. This effect could be abrogated by rapamycin suggesting that mTOR signaling may be responsible for the improvement in anemia associated with L-Leucine.
CONCLUSIONS:
Our studies support the rationale for ongoing clinical trials of L-Leucine as a therapeutic agent for DBA, and potentially for patients with del(5q) MDS.

Arch Biochem Biophys. 1955 Jul;57(1):1-12.

L-Leucine, an isoleucine antagonist in the rat[Reference: WebLink]

METHODS AND RESULTS:
An excess of dietary L-Leucine has been shown to retard the growth of rats fed low-protein diets or diets deficient in isoleucine. The addition of isoleucine to such diets overcame, to a large extent, the growth-retarding action of an excess of L-Leucine.
CONCLUSIONS:
These observations suggest that an excess of dietary L-Leucine can act as an antimetabolite of isoleucine in the rat and can thereby increase the requirement of the rat for isoleucine.

PLoS Genet. 2013;9(10):e1003857.

Stimulation of mTORC1 with L-leucine rescues defects associated with Roberts syndrome[Pubmed: 24098154 ]

Roberts syndrome (RBS) is a human disease characterized by defects in limb and craniofacial development and growth and mental retardation. RBS is caused by mutations in ESCO2, a gene which encodes an acetyltransferase for the cohesin complex. While the essential role of the cohesin complex in chromosome segregation has been well characterized, it plays additional roles in DNA damage repair, chromosome condensation, and gene expression. The developmental phenotypes of Roberts syndrome and other cohesinopathies suggest that gene expression is impaired during embryogenesis. It was previously reported that ribosomal RNA production and protein translation were impaired in immortalized RBS cells. It was speculated that cohesin binding at the rDNA was important for nucleolar form and function.
METHODS AND RESULTS:
We have explored the hypothesis that reduced ribosome function contributes to RBS in zebrafish models and human cells. Two key pathways that sense cellular stress are the p53 and mTOR pathways. We report that mTOR signaling is inhibited in human RBS cells based on the reduced phosphorylation of the downstream effectors S6K1, S6 and 4EBP1, and this correlates with p53 activation. Nucleoli, the sites of ribosome production, are highly fragmented in RBS cells. We tested the effect of inhibiting p53 or stimulating mTOR in RBS cells. The rescue provided by mTOR activation was more significant, with activation rescuing both cell division and cell death. To study this cohesinopathy in a whole animal model we used ESCO2-mutant and morphant zebrafish embryos, which have developmental defects mimicking RBS. Consistent with RBS patient cells, the ESCO2 mutant embryos show p53 activation and inhibition of the TOR pathway. Stimulation of the TOR pathway with L-Leucine rescued many developmental defects of ESCO2-mutant embryos.
CONCLUSIONS:
Our data support the idea that RBS can be attributed in part to defects in ribosome biogenesis, and stimulation of the TOR pathway has therapeutic potential.

Blood. 2012 Sep 13;120(11):2225-8.

Dietary L-leucine improves the anemia in a mouse model for Diamond-Blackfan anemia.[Pubmed: 22791294 ]

Diamond-Blackfan anemia (DBA) is a congenital erythroid hypoplasia caused by a functional haploinsufficiency of genes encoding for ribosomal proteins. Recently, a case study reported a patient who became transfusion-independent in response to treatment with the amino acid L-Leucine.
METHODS AND RESULTS:
Therefore, we have validated the therapeutic effect of L-Leucine using our recently generated mouse model for RPS19-deficient DBA. Administration of L-Leucine significantly improved the anemia in Rps19-deficient mice (19% improvement in hemoglobin concentration; 18% increase in the number of erythrocytes), increased the bone marrow cellularity, and alleviated stress hematopoiesis. Furthermore, the therapeutic response to L-Leucine appeared specific for Rps19-deficient hematopoiesis and was associated with down-regulation of p53 activity.
CONCLUSIONS:
Our study supports the rationale for clinical trials of L-Leucine as a therapeutic agent for DBA.