Clinical trial NCT01550705
Conditions: Erythropoietic Protoporphyria (EPP) X Linked Erythropoietic Protoporphyria
Keywords: Protoporphyria Porphyria Porphyrins
Sponsor: University of Utah
Start date: 2012-03-01; Completion date: 2015-12-01
Enrollment: 11 participants; Allocation: NA; Intervention model: SINGLE_GROUP

Brief summary:
In erythropoietic protoporphyria there is an accumulation of protoporphyrin IX (PPIX) in the plasma and liver. The reason it builds up is either the last step to make heme, insertion of iron into PPIX, is rate limiting or there is an increase in activity in the first step in the heme pathway.

It may be possible to decrease the amount of PPIX made and see a decrease in symptoms. The first step to make heme is the key step in the pathway and it uses vitamin B6 as a cofactor. If the investigators can limit the amount of vitamin B6 the investigators can possibly reduce the activity of this rate limiting step. With decreased activity of the enzyme it may be possible for the body to utilize all the PPIX that is made so that none builds up.

Detailed description:
Clinically, both erythropoietic protoporphyria (EPP) and X-linked EPP (XLEPP) are characterized by painful, non-blistering cutaneous photosensitivity with onset in early childhood. EPP is the most common porphyria in children and the third most common in adults (after porphyria cutanea tarda and acute intermittent porphyria). Reports of prevalence vary between 5 and 15 cases per million population.

EPP is due in most cases to decreased activity of ferrochelatase (FECH), the enzyme that catalyzes the incorporation of ferrous iron into PPIX, the final step in the production of heme. The pattern of inheritance is autosomal recessive. However, homozygosity for a FECH mutation is rare. Rather, the decreased activity is a consequence of a combination of an inherited inactivating mutation affecting one FECH allele and an intronic polymorphism that alters splicing of the other allele. The alternative splice site, when used, produces a non-functional FECH messenger ribonucleic acid (mRNA). The alternative splice site is used approximately 40% of the time. Therefore, the polymorphic allele produces approximately 60% of normal FECH activity, and for this reason, is termed hypomorphic. When the hypomorphic FECH allele is in trans with the non-functional mutant allele the result is 30% or less of the normal FECH enzyme activity. This subnormal FECH activity becomes rate-limiting, resulting in accumulation of intracellular PPIX. Although the defect is presumably expressed in all tissues, the PPIX responsible for photosensitivity derives primarily from marrow reticulocytes.

Aminolevulinic acid synthase (ALAS), the first, and rate-limiting enzyme in the heme biosynthetic pathway, catalyzes the condensation of glycine and succinyl-Coenzyme A (succinyl-CoA) to form aminolevulinic acid (ALA), and requires pyridoxal 5'-phosphate as a cofactor. ALAS in mammalian cells is localized to the mitochondrial matrix. The enzyme is synthesized as a precursor protein in the cytosol and transported into mitochondria. Two separate ALA synthase genes encode housekeeping (tissue-nonspecific) and erythroid specific forms of the enzyme (ALAS1 and ALAS2, respectively). The gene for human ALAS1 is on 3p.21 and the locus for ALAS2 the X-chromosome, at Xp11.2.

The two forms of ALAS are differentially regulated, ALAS1 is a housekeeping gene expressed in all cells and ALAS2 is driven by erythroid specific transcription factors GATA1 and NF-E2. Additionally, ALAS2 mRNA contains an iron-responsive element (IRE) in its 5'-untranslated region, similar to mRNAs encoding ferritin and the transferrin receptor (in which the IRE is in the gene's 3' UTR). Gel retardation analysis showed that the iron-responsive element in ALAS2 mRNA is functional \[as evidenced by binding to iron regulator protein 2 (IRP2)\], indicating that translation of the erythroid-specific mRNA is directly linked to the availability of iron and heme in erythroid cells. In this case, when intracellular iron concentration is relatively high, it is available for binding to IRP2, a process that enhances ubiquitin-mediated degradation of IRP2. Under these conditions, IRP2 is unavailable for binding to the IRE element in ALAS2, clearing the message for efficient translation. Conversely, when intracellular iron is relatively low, IRP2 degradation is restricted, making the protein available for binding to the IRE and thereby blocking translation of ALAS2 mRNA.

Recently, a variant form of EPP, inherited in an X-linked pattern (XLEPP), was shown to be due to an ALAS2 gain-of-function mutation in exon 11. The mutation results in a truncated form of the protein that has supranormal specific activity as a result of less constrained enzyme-substrate interactions, resulting in overproduction of PPIX. This situation is in contrast to EPP with mutated FECH in which PPIX accumulates because of deficient heme formation.

ALAS1 and 2 use pyridoxal phosphate (PLP) as a cofactor. PLP is a modified form of vitamin B6. It has been shown that PLP complexes with isoniazid depleting the cofactor. This PLP depletion has been one of the causes of sideroblastic anemia.

The investigators will test the hypothesis that depletion of PLP will lead to decreased activity of ALAS.

Arms: arm 1: Subjects will receive isoniazid daily for 2 months. Subjects will be seen every 2 weeks to obtain lab samples and health check.
Interventions: intervention 1: Isoniazid
Intervention descriptions: intervention 1: Isoniazid 5 mg/Kg up to 300 mg per day. Oral tablets. 2 months.

Protocol documents: