PMID- 32582668
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20200928
IS  - 2296-4185 (Print)
IS  - 2296-4185 (Electronic)
IS  - 2296-4185 (Linking)
VI  - 8
DP  - 2020
TI  - Challenges and Pitfalls in the Engineering of Human Interleukin 22 (hIL-22) 
      Secreting Lactobacillus reuteri.
PG  - 543
LID - 10.3389/fbioe.2020.00543 [doi]
LID - 543
AB  - Engineered microbes for the delivery of intestinally directed therapeutics is a 
      promising avenue for the treatment of various intestinal diseases including 
      inflammatory bowel disease (IBD) and intestinal graft vs. host disease (GVHD). 
      This modality of treatment would allow for the targeted delivery of therapeutics 
      to the site of inflammation or disease while minimizing the systemic side effects 
      that often accompany treatment of these pathologies. Here, we show the challenges 
      encountered and overcome in successfully engineering Lactobacillus reuteri to 
      secrete high levels of biologically active human interleukin 22 (hIL-22). Initial 
      hIL-22 constructs secreted high levels of hIL-22, however we found the majority 
      of hIL-22 was cleaved and not biologically active. Several strategies were 
      explored to improve the production of intact hIL-22, with the optimization of the 
      signal sequence for peptide secretion having the most impact of production of 
      intact hIL-22. This resulted in L. reuteri secreting high concentrations (up to 
      700 ng/mL) of hIL-22. Bioactivity of hIL-22 was confirmed by the secretion of 
      interleukin 10 (IL-10) from the colon cancer derived epithelial cell line Colo205 
      and the secretion of Regenerating islet-derived protein 3 alpha (Reg3alpha) from 
      human jejunal enteroids. The secretion of bioactive hIL-22 imposed a significant 
      cost for L. reuteri as bacterial growth was significantly impaired upon 
      induction. Future challenges and optimization strategies for the delivery of 
      hIL-22 to the human intestinal tract are discussed.
CI  - Copyright (c) 2020 Ortiz-Velez, Goodwin, Schaefer and Britton.
FAU - Ortiz-Velez, Laura
AU  - Ortiz-Velez L
AD  - Department of Molecular Virology and Microbiology, Baylor College of Medicine, 
      Houston, TX, United States.
FAU - Goodwin, Annie
AU  - Goodwin A
AD  - Department of Molecular Virology and Microbiology, Baylor College of Medicine, 
      Houston, TX, United States.
AD  - Section of Pediatric Gastroenterology, Department of Pediatrics, Baylor College 
      of Medicine, Texas Children's Hospital, Houston, TX, United States.
FAU - Schaefer, Laura
AU  - Schaefer L
AD  - Department of Molecular Virology and Microbiology, Baylor College of Medicine, 
      Houston, TX, United States.
FAU - Britton, Robert A
AU  - Britton RA
AD  - Department of Molecular Virology and Microbiology, Baylor College of Medicine, 
      Houston, TX, United States.
AD  - Alkek Center for Metagenomics and Microbiome Research, Baylor College of 
      Medicine, Houston, TX, United States.
LA  - eng
GR  - P01 CA039542/CA/NCI NIH HHS/United States
GR  - T32 DK007664/DK/NIDDK NIH HHS/United States
PT  - Journal Article
DEP - 20200605
PL  - Switzerland
TA  - Front Bioeng Biotechnol
JT  - Frontiers in bioengineering and biotechnology
JID - 101632513
PMC - PMC7289926
OTO - NOTNLM
OT  - IL-22
OT  - lactobacilus
OT  - probiotic
OT  - secretion
OT  - therapeutic delivery
EDAT- 2020/06/26 06:00
MHDA- 2020/06/26 06:01
PMCR- 2020/01/01
CRDT- 2020/06/26 06:00
PHST- 2019/09/25 00:00 [received]
PHST- 2020/05/05 00:00 [accepted]
PHST- 2020/06/26 06:00 [entrez]
PHST- 2020/06/26 06:00 [pubmed]
PHST- 2020/06/26 06:01 [medline]
PHST- 2020/01/01 00:00 [pmc-release]
AID - 10.3389/fbioe.2020.00543 [doi]
PST - epublish
SO  - Front Bioeng Biotechnol. 2020 Jun 5;8:543. doi: 10.3389/fbioe.2020.00543. 
      eCollection 2020.