Low cost industrial production of coagulation factor IX bioencapsulated in lettuce cells for oral tolerance induction in hemophilia B.

Abstract

Antibodies (inhibitors) developed by hemophilia B patients against coagulation factor IX (FIX) are challenging to eliminate because of anaphylaxis or nephrotic syndrome after continued infusion. To address this urgent unmet medical need, FIX fused with a transmucosal carrier (CTB) was produced in a commercial lettuce (Simpson Elite) cultivar using species specific chloroplast vectors regulated by endogenous psbA sequences. CTB-FIX (∼1 mg/g) in lyophilized cells was stable with proper folding, disulfide bonds and pentamer assembly when stored ∼2 years at ambient temperature. Feeding lettuce cells to hemophilia B mice delivered CTB-FIX efficiently to the gut immune system, induced LAP(+) regulatory T cells and suppressed inhibitor/IgE formation and anaphylaxis against FIX. Lyophilized cells enabled 10-fold dose escalation studies and successful induction of oral tolerance was observed in all tested doses. Induction of tolerance in such a broad dose range should enable oral delivery to patients of different age groups and diverse genetic background. Using Fraunhofer cGMP hydroponic system, ∼870 kg fresh or 43.5 kg dry weight can be harvested per 1000 ft(2) per annum yielding 24,000-36,000 doses for 20-kg pediatric patients, enabling first commercial development of an oral drug, addressing prohibitively expensive purification, cold storage/transportation and short shelf life of current protein drugs.

• Authors: Jin Su, Liqing Zhu, Alexandra Sherman, Xiaomei Wang, Shina Lin, Aditya Kamesh, Joey H Norikane, Stephen J Streatfield, Roland W Herzog, Henry Daniell
• Journal: Biomaterials (Biomaterials) Vol. 70 Pg. 84-93 (Nov 2015) ISSN: 1878-5905 [Electronic] Netherlands
• PMID: 26302233 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

Chemical References

• Antigens
• Capsules
• Immunoglobulin E
• Factor IX

Topics

• Administration, Oral
• Animals
• Antigens (metabolism)
• Biomass
• Capsules
• Chloroplasts (metabolism)
• Costs and Cost Analysis
• Factor IX (administration & dosage, economics, therapeutic use)
• Freeze Drying
• Genetic Vectors (metabolism)
• Hemophilia B (drug therapy)
• Immune Tolerance
• Immunoglobulin E (metabolism)
• Industry
• Intestine, Small (metabolism)
• Lactuca (cytology)
• Mice
• Plant Leaves (metabolism)
• Protein Folding
• Protein Stability