Preclinical development of plant-based oral immune modulatory therapy for haemophilia B.
Preclinical development of plant-based oral immune modulatory therapy for haemophilia B.
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DOI：
10.1111/pbi.13608
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发表时间：
2021-10
影响因子：
13.8
通讯作者：
Daniell H
中科院分区：
文献类型：
--
作者：
Srinivasan A;Herzog RW;Khan I;Sherman A;Bertolini T;Wynn T;Daniell H
抗药物抗体 (ADA) 的形成是治疗 X 连锁出血性疾病 B 型血友病（凝血因子 IX、FIX 缺乏）的主要并发症。目前的临床免疫耐受方案通常由于 FIX 过敏反应等并发症而无效。基于植物的口服耐受诱导可能会解决这个问题，最近监管部门首次批准口服植物细胞来治疗花生过敏就说明了这一点。我们之前的研究表明，口服表达 FIX 的植物细胞与叶绿体中的跨粘膜载体 CTB（霍乱毒素亚基 B）融合，可以抑制 B 型血友病动物的 ADA。我们在这里报告了第一个在不存在抗生素抗性基因的情况下表达凝血因子的生菜转质体系。 T1 和 T2 代无标记植物中均保持了 CTB-FIX 基因的稳定整合和同质性（每个细胞中约 10,000 个拷贝的转化）。 T1 和 T2 无标记植物的冻干叶子中的 CTB-FIX 表达为 1.0–1.5 mg/g 干重，证实标记切除不会影响抗原水平。给 Sprague Dawley 大鼠口服 0.25、1 或 2.5 mg/kg 的 CTB-FIX 不会产生明显的不良反应或毒性。大鼠单次口服给药的未观察到不良反应水平 (NOAEL) 至少为 2.5 mg/kg。比格犬口服 0.3 或 1.47 mg/kg 的 CTB-FIX，无论是混合在食物中还是作为口服混悬液，均不会产生任何可观察到的毒性。这些毒理学研究应有助于提交监管批准文件和对 B 型血友病患者进行评估。
Anti-drug antibody (ADA) formation is a major complication in treatment of the X-linked bleeding disorder hemophilia B (deficiency in coagulation factor IX, FIX). Current clinical immune tolerance protocols are often not effective due to complications such as anaphylactic reactions against FIX. Plant-based oral tolerance induction may address this problem, as illustrated by the recent first regulatory approval of orally delivered plant cells to treat peanut allergy. Our previous studies showed that oral delivery of plant cells expressing FIX fused to the transmucosal carrier CTB (cholera toxin subunit B) in chloroplasts suppressed ADA in animals with hemophilia B. We report here creation of the first lettuce transplastomic lines expressing a coagulation factor, in the absence of antibiotic resistance gene. Stable integration of the CTB-FIX gene and homoplasmy (transformation of ~10,000 copies in each cell) were maintained in both T1 and T2 generation marker-free plants. CTB-FIX expression in lyophilized leaves of T1 and T2 marker-free plants was 1.0–1.5 mg/g dry weight, confirming that the marker excision did not affect antigen levels. Oral administration of CTB-FIX to Sprague Dawley rats at 0.25, 1 or 2.5 mg/kg did not produce overt adverse effects or toxicity. The no-observed-adverse-effect-level (NOAEL) is at least 2.5 mg/kg for a single oral administration in rats. Oral administration of CTB-FIX at 0.3 or 1.47 mg/kg either mixed in food or as an oral suspension to Beagle dogs did not produce any observable toxicity. These toxicology studies should facilitate filing of regulatory approval documents and evaluation in hemophilia B patients.
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DOI：
10.1111/hae.12229
发表时间：
2014-01
期刊：
Haemophilia : the official journal of the World Federation of Hemophilia
影响因子：
--
作者：
Puetz J;Soucie JM;Kempton CL;Monahan PE;Hemophilia Treatment Center Network (HTCN) Investigators
通讯作者：
Hemophilia Treatment Center Network (HTCN) Investigators
影响因子：
3.6
作者：
Bone Calvo, Javier;Clavero Adell, Marcos;Aliaga Mazas, Yolanda
通讯作者：
Aliaga Mazas, Yolanda
DOI：
10.1182/asheducation-2017.1.605
发表时间：
2017-12-01
影响因子：
3
作者：
Ragni, Margaret V.
通讯作者：
Ragni, Margaret V.
DOI：
10.1016/j.ymthe.2016.11.009
发表时间：
2017-02-01
期刊：
Molecular therapy : the journal of the American Society of Gene Therapy
影响因子：
--
作者：
Herzog RW;Nichols TC;Su J;Zhang B;Sherman A;Merricks EP;Raymer R;Perrin GQ;Häger M;Wiinberg B;Daniell H
通讯作者：
Daniell H
影响因子：
4.3
作者：
Bertolini TB;Biswas M;Terhorst C;Daniell H;Herzog RW;Piñeros AR
通讯作者：
Piñeros AR