Seed-Specific Expression of Apolipoprotein A-IMilano Dimer in Engineered Rice Lines
Received date: 2023-01-21
Accepted date: 2023-06-19
Online published: 2023-06-29
Apolipoprotein A-IMilano (ApoA-IM) has been shown to significantly reduce coronary atherosclerotic plaques. However, the preparation of cost-effective pharmaceutical formulations of ApoA-IM is limited by the high cost and difficulty of purifying the protein and producing the highly effective dimeric form. The aim of this study was to create an expression cassette that specifically drives the expression of dimeric ApoA-IM in the protein bodies of rice seeds. The ApoA-IM protein under control of the 13 kDa prolamin promoter is expressed exclusively in its dimeric form within the seeds, and immunocytochemical and immunogold analyses confirmed its expression in different caryopsis tissue such as seed coat, aleurone cell and endosperm, particularly in amyloplast and storage vacuoles. A plant-based ApoA-IM production system offered numerous advantages over current production systems, including the direct production of the most therapeutically effective dimeric ApoA-IM forms, long-term protein storage in seeds, and ease of protein production by simply growing plants. Therefore, seeds had the potential to serve as a cost-effective source of therapeutic ApoA-IM.
Serena Reggi, Elisabetta Onelli, Alessandra Moscatelli, Nadia Stroppa, Matteo Dell’Anno, Kiril Perfanov, Luciana Rossi . Seed-Specific Expression of Apolipoprotein A-IMilano Dimer in Engineered Rice Lines[J]. Rice Science, 2023 , 30(6) : 587 -597 . DOI: 10.1016/j.rsci.2023.09.001
| [1] | Alexander E T, Tanaka M, Kono M, Saito H, Rader D J, Phillips M C. 2009. Structural and functional consequences of the Milano mutation (R173C) in human apolipoprotein A-I. J Lipid Res, 50(7): 1409-1419. |
| [2] | Baslam M, Oikawa K, Kitajima-Koga A, Kaneko K, Mitsui T. 2016. Golgi-to-plastid trafficking of proteins through secretory pathway: Insights into vesicle-mediated import toward the plastids. Plant Signal Behav, 11(9): e1221558. |
| [3] | Ben-Aicha S, Badimon L, Vilahur G. 2020. Advances in HDL: Much more than lipid transporters. Int J Mol Sci, 21(3): 732. |
| [4] | Chiesa G, Sirtori C R. 2003. Apolipoprotein A-IMilano: Current perspectives. Curr Opin Lipidol, 14(2): 159-163. |
| [5] | Chyu K Y, Shah P K. 2015. HDL/ApoA-1 infusion and ApoA-1 gene therapy in atherosclerosis. Front Pharmacol, 6: 187. |
| [6] | Doyle J J, Doyle J L. 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull, 19(1): 11-15. |
| [7] | Emancipator K, Csako G, Elin R J. 1992. In vitro inactivation of bacterial endotoxin by human lipoproteins and apolipoproteins. Infect Immun, 60(2): 596-601. |
| [8] | Gelvin S B. 2003. Agrobacterium-mediated plant transformation: The biology behind the “gene-jockeying” tool. Microbiol Mol Biol Rev, 67(1): 16-37. |
| [9] | Gogonea V, Gerstenecker G S, Wu Z P, Lee X, Topbas C, Wagner M A, Tallant T C, Smith J D, Callow P, Pipich V, Malet H, Schoehn G, DiDonato J A, Hazen S L. 2013. The low-resolution structure of nHDL reconstituted with DMPC with and without cholesterol reveals a mechanism for particle expansion. J Lipid Res, 54(4): 966-983. |
| [10] | Gunasekaran B, Gothandam K M. 2020. A review on edible vaccines and their prospects. Braz J Med Biol Res, 53(2): e8749. |
| [11] | Gupta R, Brunak S. 2002. Prediction of glycosylation across the human proteome and the correlation to protein function. Pac Symp Biocomput, 7: 310-322. |
| [12] | Hiei Y, Ohta S, Komari T, Kumashiro T. 1994. Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J, 6(2): 271-282. |
| [13] | Jain N, Vergish S, Khurana J P. 2018. Validation of house-keeping genes for normalization of gene expression data during diurnal/ circadian studies in rice by RT-qPCR. Sci Rep, 8: 3203. |
| [14] | Kim S J, Brandizzi F. 2014. The plant secretory pathway: An essential factory for building the plant cell wall. Plant Cell Physiol, 55(4): 687-693. |
| [15] | Kurup V M, Thomas J. 2020. Edible vaccines: Promises and challenges. Mol Biotechnol, 62(2): 79-90. |
| [16] | Laemmli U K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227: 680-685. |
| [17] | Livak K J, Schmittgen T D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods, 25(4): 402-408. |
| [18] | Lorenzetti R, Sidoli A, Palomba R, Monaco L, Martineau D, Lappi D A, Soria M. 1986. Expression of the human apolipoprotein AI gene fused to the E. coligene for β-galactosidase. FEBS Lett, 194(2): 347-350. |
| [19] | Mamat U, Wilke K, Bramhill D, Schromm A B, Lindner B, Kohl T A, Corchero J L, Villaverde A, Schaffer L, Head S R, Souvignier C, Meredith T C, Woodard R W. 2015. Detoxifying Escherichia coli for endotoxin-free production of recombinant proteins. Microb Cell Fact, 14: 57. |
| [20] | Moscatelli A, Gagliardi A, Maneta-Peyret L, Bini L, Stroppa N, Onelli E, Landi C, Scali M, Idilli A I, Moreau P. 2015. Characterisation of detergent-insoluble membranes in pollen tubes of Nicotiana tabacum (L.). Biol Open, 4(3): 378-399. |
| [21] | Muench D G, Ogawa M, Okita T W. 1999. The prolamins of rice. In: Shewry P R, Casey R. Seed Proteins. Dordrecht, the Netherlands: Springer: 93-108. |
| [22] | Nissen S E, Tsunoda T, Tuzcu E M, Schoenhagen P, Cooper C J, Yasin M, Eaton G M, Lauer M A, Sheldon W S, Grines C L, Halpern S, Crowe T, Blankenship J C, Kerensky R. 2003. Effect of recombinant ApoA-I Milano on coronary atherosclerosis in patients with acute coronary syndromes: A randomized controlled trial. JAMA, 290(17): 2292-2300. |
| [23] | Nykiforuk C L, Shen Y, Murray E W, Boothe J G, Busseuil D, Rhéaume E, Tardif J C, Reid A, Moloney M M. 2011. Expression and recovery of biologically active recombinant Apolipoprotein AIMilano from transgenic safflower (Carthamus tinctorius) seeds. Plant Biotechnol J, 9(2): 250-263. |
| [24] | Oluwayelu D O, Adebiyi A I. 2016. Plantibodies in human and animal health: A review. Afr Health Sci, 16(2): 640-645. |
| [25] | Onelli E, Moscatelli A, Gagliardi A, Zaninelli M, Bini L, Baldi A, Caccianiga M, Reggi S, Rossi L. 2017. Retarded germination of Nicotiana tabacum seeds following insertion of exogenous DNA mimics the seed persistent behavior. PLoS One, 12(12): e0187929. |
| [26] | Petrlova J, Dalla-Riva J, Mörgelin M, Lindahl M, Krupinska E, Stenkula K G, Voss J C, Lagerstedt J O. 2014. Secondary structure changes in ApoA-I Milano (R173C) are not accompanied by a decrease in protein stability or solubility. PLoS One, 9(4): e96150. |
| [27] | Qu L Q, Takaiwa F. 2004. Evaluation of tissue specificity and expression strength of rice seed component gene promoters in transgenic rice. Plant Biotechnol J, 2(2): 113-125. |
| [28] | Romano G, Reggi S, Kutryb-Zajac B, Facoetti A, Chisci E, Pettinato M, Giuffrè M R, Vecchio F, Leoni S, De Giorgi M, Avezza F, Cadamuro M, Crippa L, Leone B E, Lavitrano M, Rivolta I, Barisani D, Smolenski R T, Giovannoni R. 2018. APOA-1Milano muteins, orally delivered via genetically modified rice, show anti-atherogenic and anti-inflammatory properties in vitro and in Apoe-/- atherosclerotic mice. Int J Cardiol, 271: 233-239. |
| [29] | Rosano G L, Ceccarelli E A. 2014. Recombinant protein expression in Escherichia coli: Advances and challenges. Front Microbiol, 5: 172. |
| [30] | Rossi L, Dell’Orto V, Vagni S, Sala V, Reggi S, Baldi A. 2014a. Protective effect of oral administration of transgenic tobacco seeds against verocytotoxic Escherichia coli strain in piglets. Vet Res Commun, 38(1): 39-49. |
| [31] | Rossi L, Pinotti L, Agazzi A, Dell’Orto V, Baldi A. 2014b. Plant bioreactors for the antigenic hook-associated flgK protein expression. Ital J Animal Sci, 13(1): 2939. |
| [32] | Ryan R O, Forte T M, Oda M N. 2003. Optimized bacterial expression of human apolipoprotein A-I. Protein Expr Purif, 27(1): 98-103. |
| [33] | Saito Y, Shigemitsu T, Yamasaki R, Sasou A, Goto F, Kishida K, Kuroda M, Tanaka K, Morita S, Satoh S, Masumura T. 2012. Formation mechanism of the internal structure of type I protein bodies in rice endosperm: Relationship between the localization of prolamin species and the expression of individual genes. Plant J, 70(6): 1043-1055. |
| [34] | Schmidt H H J, Genschel J, Haas R, Büttner C, Manns M P. 1997. Expression and purification of recombinant human apolipoprotein A-I in Chinese Hamster ovary cells. Protein Expr Purif, 10(2): 226-236. |
| [35] | Sinha P, Poland J, Schnölzer M, Rabilloud T. 2001. A new silver staining apparatus and procedure for matrix-assisted laser desorption/ionization-time of flight analysis of proteins after two-dimensional electrophoresis. Proteomics, 1(7): 835-840. |
| [36] | Sirtori C R, Ruscica M, Calabresi L, Chiesa G, Giovannoni R, Badimon J J. 2019. HDL therapy today: From atherosclerosis, to stent compatibility to heart failure. Ann Med, 51(7/8): 345-359. |
| [37] | Smith J D. 2010. Apolipoprotein AI and its mimetics for the treatment of atherosclerosis. Curr Opin Invest Drugs, 11(9): 989-996. |
| [38] | Stoffel W. 1984. Synthesis, transport, and processing of apolipoproteins of high density lipoproteins. J Lipid Res, 25(13): 1586-1592. |
| [39] | Streatfield S J. 2007. Approaches to achieve high-level heterologous protein production in plants. Plant Biotechnol J, 5(1): 2-15. |
| [40] | Takaiwa F, Kikuchi S, Oono K. 1987. A rice glutelin gene family: A major type of glutelin mRNAs can be divided into two classes. Mol Gen Genet, 208(1/2): 15-22. |
| [41] | Takaiwa F, Wakasa Y, Hayashi S, Kawakatsu T. 2017. An overview on the strategies to exploit rice endosperm as production platform for biopharmaceuticals. Plant Sci, 263: 201-209. |
| [42] | Tanaka K, Sugimoto T, Ogawa M, Kasai Z. 1980. Isolation and characterization of two types of protein bodies in the rice endosperm. Agric Biol Chem, 44(7): 1633-1639. |
| [43] | Tardif J C, Grégoire J, L’Allier P L, Ibrahim R, Lespérance J, Heinonen T M, Kouz S, Berry C, Basser R, Lavoie M A, Guertin M C, Rodés-Cabau J. 2007. Effects of reconstituted high-density lipoprotein infusions on coronary atherosclerosis: A randomized controlled trial. JAMA, 297: 1675-1682. |
| [44] | Torres E, Gonzalez-Melendi P, Sto?ger E, Shaw P, Twyman R M, Nicholson L, Vaquero C, Fischer R, Christou P, Perrin Y. 2001. Native and artificial reticuloplasmins co-accumulate in distinct domains of the endoplasmic reticulum and in post-endoplasmic reticulum compartments. Plant Physiol, 127(3): 1212-1223. |
| [45] | Watts D, MacBeath J R. 2001. Automated fluorescent DNA sequencing on the ABI PRISM 310 Genetic Analyzer. Methods Mol Biol, 167: 153-170. |
| [46] | Zhu Q L, Tan J T, Liu Y G. 2022. Molecular farming using transgenic rice endosperm. Trends Biotechnol, 40(10): 1248-1260. |
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