mRNA-based therapy in a rabbit model of variegate porphyria offers new insights into the pathogenesis of acute attacks

Jericó, Daniel; Córdoba, Karol M.; Jiang, Lei; Morán, María; Sampedro, Ana; Alegre, Manuel; Collantes, María; Santamaría, Eva; Alegre, Estíbaliz; Culerier, Corinne; Mendoza, Ander Estella-Hermoso de; Oyarzábal, Julen; Martı́n, Miguel; Peñuelas, Iván; Ávila, Matías A.; Gouya, Laurent; Martini, Paolo G.V.; Fontanellas, Antonio

doi:10.1016/j.omtn.2021.05.010

Cited by 14 publications

(14 citation statements)

References 57 publications

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“…This corresponds to the phenotype observed in PPO-KO cells that reveal compromised oxygen consumption, decreased CIII and CIV quantities and aberrant assembly of CI n CIII n (super)complexes. It is interesting to note that the link between dysregulated heme levels accompanying VP and compromised function of OXPHOS complexes, mainly CIV, has been recently observed using a rabbit model of induced VP in vivo 58 , corroborating thus findings reported here. In their paper, Jerico et al showed that the insufficient efficiency of derivatization of protoheme to heme a might be responsible for the decreased levels and the compromised function of CIV.…”

Section: Discussionsupporting

confidence: 90%

Generation and characterization of human U-2 OS cell lines with the CRISPR/Cas9-edited protoporphyrinogen oxidase IX gene

Nováková

Milošević

Kutil

et al. 2022

Sci Rep

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In humans, disruptions in the heme biosynthetic pathway are associated with various types of porphyrias, including variegate porphyria that results from the decreased activity of protoporphyrinogen oxidase IX (PPO; E.C.1.3.3.4), the enzyme catalyzing the penultimate step of the heme biosynthesis. Here we report the generation and characterization of human cell lines, in which PPO was inactivated using the CRISPR/Cas9 system. The PPO knock-out (PPO-KO) cell lines are viable with the normal proliferation rate and show massive accumulation of protoporphyrinogen IX, the PPO substrate. Observed low heme levels trigger a decrease in the amount of functional heme containing respiratory complexes III and IV and overall reduced oxygen consumption rates. Untargeted proteomics further revealed dysregulation of 22 cellular proteins, including strong upregulation of 5-aminolevulinic acid synthase, the major regulatory protein of the heme biosynthesis, as well as additional ten targets with unknown association to heme metabolism. Importantly, knock-in of PPO into PPO-KO cells rescued their wild-type phenotype, confirming the specificity of our model. Overall, our model system exploiting a non-erythroid human U-2 OS cell line reveals physiological consequences of the PPO ablation at the cellular level and can serve as a tool to study various aspects of dysregulated heme metabolism associated with variegate porphyria.

show abstract

Section: Discussionsupporting

confidence: 90%

Generation and characterization of human U-2 OS cell lines with the CRISPR/Cas9-edited protoporphyrinogen oxidase IX gene

Nováková

Milošević

Kutil

et al. 2022

Sci Rep

View full text Add to dashboard Cite

show abstract

“…This corresponds to the phenotype observed in PPO-KO cells that reveal compromised oxygen consumption, decreased CIII and CIV quantities and aberrant assembly of CI n CIII n (super)complexes. It is interesting to note that the link between dysregulated heme levels accompanying VP and compromised function of OXPHOS complexes, mainly CIV, has been recently observed using a rabbit model of induced VP in vivo 55 , corroborating thus ndings reported here. In their paper, Jerico et al showed that the insu cient e ciency of derivatization of protoheme to heme a might be responsible for the decreased levels and the compromised function of CIV.…”

Section: Discussionsupporting

confidence: 90%

Generation and characterization of human U-2 OS cell lines with the CRISPR/Cas9-edited protoporphyrinogen oxidase IX gene

Nováková

Milošević

Kutil

et al. 2022

Preprint

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In humans, disruptions in the heme biosynthetic pathway are associated with various types of porphyrias, including variegate porphyria that results from the decreased activity of protoporphyrinogen oxidase IX (PPO; E.C.1.3.3.4), the enzyme catalyzing the penultimate step of the heme biosynthesis. Here we report the generation and characterization of human cell lines, in which PPO was inactivated using the CRISPR/Cas9 system. The PPO knock-out (PPO-KO) cell lines are viable with the normal proliferation rate and show massive accumulation of protoporphyrinogen IX, the PPO substrate. Observed low heme levels trigger a decrease in the amount of functional heme containing respiratory complexes III and IV and overall reduced oxygen consumption rates. Untargeted proteomics further revealed dysregulation of 22 cellular proteins, including strong upregulation of 5-aminolevulinic acid synthase, the major regulatory protein of the heme biosynthesis, as well as additional ten targets with unknown association to heme metabolism. Importantly, knock-in of PPO into PPO-KO cells rescued their wild-type phenotype, confirming the specificity of our model. Overall, our model system reveals physiological consequences of the PPO ablation at the cellular level and can serve as a tool to study various aspects of dysregulated heme metabolism associated with variegate porphyria.

show abstract

“…mRNA-based therapy is an emerging approach with great potential for the treatment of liver diseases [159]. Indeed, systemic administration of engineered mRNAs formulated in lipid nanoparticles (mRNA-LNPs) shielded from humoral and cellular immunity showed high efficacy for the treatment of experimental hepatic porphyria in mouse and rabbit models [160,161]. When these mRNA-LNPs were tested in nonhuman primates the hepatic parenchyma was robustly transduced, and no signs of toxicity nor immune response were observed [160].…”

Section: Is It Possible To Recover Hepatocellular Identity In a Chron...mentioning

confidence: 99%

Loss of liver function in chronic liver disease: An identity crisis

Berasain¹,

Arechederra²,

Argemí³

et al. 2023

Journal of Hepatology

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mRNA-based therapy in a rabbit model of variegate porphyria offers new insights into the pathogenesis of acute attacks

Cited by 14 publications

References 57 publications

Generation and characterization of human U-2 OS cell lines with the CRISPR/Cas9-edited protoporphyrinogen oxidase IX gene

Generation and characterization of human U-2 OS cell lines with the CRISPR/Cas9-edited protoporphyrinogen oxidase IX gene

Generation and characterization of human U-2 OS cell lines with the CRISPR/Cas9-edited protoporphyrinogen oxidase IX gene

Loss of liver function in chronic liver disease: An identity crisis

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