Top Oguz, Geisen Ulrich, Decker Eva L, Reski Ralf
Plant Biotechnology, Faculty of Biology, University of Freiburg, Freiburg im Breisgau, Germany.
Spemann Graduate School of Biology and Medicine, University of Freiburg, Freiburg im Breisgau, Germany.
Front Plant Sci. 2019 Mar 7;10:261. doi: 10.3389/fpls.2019.00261. eCollection 2019.
The use of plants as production platforms for pharmaceutical proteins has been on the rise for the past two decades. The first marketed plant-made pharmaceutical, taliglucerase alfa against Gaucher's disease produced in carrot cells by Pfizer/Protalix Biotherapeutics, was approved by the US Food and Drug Administration (FDA) in 2012. The advantages of plant systems are low cost and highly scalable biomass production compared to the fermentation systems, safety compared with other expression systems, as plant-based systems do not produce endotoxins, and the ability to perform complex eukaryotic post-translational modifications, e.g., -glycosylation that can be further engineered to achieve humanized -glycan structures. Although bleeding disorders affect only a small portion of the world population, costs of clotting factor concentrates impose a high financial burden on patients and healthcare systems. The majority of patients, ∼75% in the case of hemophilia, have no access to an adequate treatment. The necessity of large-scale and less expensive production of human blood coagulation factors, particularly factors associated with rare bleeding disorders, may be an important area for plant-based systems, as coagulation factors do not fit into the industry-favored production models. In this review, we explore previous studies on recombinant production of coagulation Factor II, VIII, IX, and XIII in different plant species. Production of bioactive FII and FIX in plants was not achieved yet due to complex post-translational modifications, including vitamin K-dependent γ-carboxylation and propeptide removal. Although plant-made FVIII and FXIII showed specific activities, there are no follow-up studies like pre-clinical/clinical trials. Significant progress has been achieved in oral delivery of bioencapsulated FVIII and FIX to induce immune tolerance in murine models of hemophilia A and B, resp. Potential strategies to overcome bottlenecks in the production systems are also addressed in this review.
在过去二十年中，利用植物作为药用蛋白质的生产平台的情况一直在增加。首个上市的植物源制药产品是辉瑞/Protagen生物治疗公司在胡萝卜细胞中生产的用于治疗戈谢病的他利糖苷酶α，于2012年获得美国食品药品监督管理局（FDA）批准。与发酵系统相比，植物系统的优势在于成本低且生物质产量具有高度可扩展性，与其他表达系统相比安全性高，因为基于植物的系统不会产生内毒素，并且能够进行复杂的真核生物翻译后修饰，例如可以进一步工程化以实现人源化聚糖结构的N-糖基化。尽管出血性疾病仅影响世界人口的一小部分，但凝血因子浓缩物的成本给患者和医疗保健系统带来了沉重的经济负担。大多数患者，如血友病患者中约75%，无法获得充分的治疗。大规模且低成本生产人凝血因子，特别是与罕见出血性疾病相关的因子的必要性，可能是基于植物的系统的一个重要领域，因为凝血因子不符合行业青睐的生产模式。在本综述中，我们探讨了先前关于在不同植物物种中重组生产凝血因子II、VIII、IX和XIII的研究。由于复杂的翻译后修饰，包括维生素K依赖性γ-羧化和前肽去除，尚未在植物中实现生物活性FII和FIX的生产。尽管植物源FVIII和FXIII显示出特定活性，但没有像临床前/临床试验这样的后续研究。在将生物包封的FVIII和FIX口服递送至血友病A和B的小鼠模型中以诱导免疫耐受方面分别取得了重大进展。本综述还讨论了克服生产系统瓶颈的潜在策略。
