PMID- 28213849 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20200930 IS - 2192-8304 (Print) IS - 2192-8312 (Electronic) IS - 2192-8304 (Linking) VI - 36 DP - 2017 TI - Widespread Expression of a Membrane-Tethered Version of the Soluble Lysosomal Enzyme Palmitoyl Protein Thioesterase-1. PG - 85-92 LID - 10.1007/8904_2017_1 [doi] AB - "Cross-correction," the transfer of soluble lysosomal enzymes between neighboring cells, forms the foundation for therapeutics of lysosomal storage disorders (LSDs). However, "cross-correction" poses a significant barrier to studying the role of specific cell types in LSD pathogenesis. By expressing the native enzyme in only one cell type, neighboring cell types are invariably corrected. In this study, we present a strategy to limit "cross-correction" of palmitoyl-protein thioesterase-1(PPT1), a lysosomal hydrolase deficient in Infantile Neuronal Ceroid Lipofuscinosis (INCL, Infantile Batten disease) to the lysosomal membrane via the C-terminus of lysosomal associated membrane protein-1 (LAMP1). Tethering PPT1 to the lysosomal membrane prevented "cross-correction" while allowing PPT1 to retain its enzymatic function and localization in vitro. A transgenic line harboring the lysosomal membrane-tethered PPT1 was then generated. We show that expression of lysosome-restricted PPT1 in vivo largely rescues the INCL biochemical, histological, and functional phenotype. These data suggest that lysosomal tethering of PPT1 via the C-terminus of LAMP1 is a viable strategy and that this general approach can be used to study the role of specific cell types in INCL pathogenesis, as well as other LSDs. Ultimately, understanding the role of specific cell types in the disease progression of LSDs will help guide the development of more targeted therapeutics. One Sentence Synopsis: Tethering PPT1 to the lysosomal membrane is a viable strategy to prevent "cross-correction" and will allow for the study of specific cellular contributions in INCL pathogenesis. FAU - Shyng, Charles AU - Shyng C AD - Department of Internal Medicine, Washington University School of Medicine, Campus Box 8007, 660 S. Euclid Ave, St. Louis, MO, 63110, USA. FAU - Macauley, Shannon L AU - Macauley SL AD - Department of Neurology, Washington University School of Medicine, St. Louis, MO, 63110, USA. FAU - Dearborn, Joshua T AU - Dearborn JT AD - Department of Internal Medicine, Washington University School of Medicine, Campus Box 8007, 660 S. Euclid Ave, St. Louis, MO, 63110, USA. FAU - Sands, Mark S AU - Sands MS AD - Department of Internal Medicine, Washington University School of Medicine, Campus Box 8007, 660 S. Euclid Ave, St. Louis, MO, 63110, USA. mssands@wustl.edu. AD - Department of Genetics, Washington University School of Medicine, St. Louis, MO, 63110, USA. mssands@wustl.edu. AD - Hope Center for Neurological Disorders, Washington University School of Medicine, St. Louis, MO, 63110, USA. mssands@wustl.edu. LA - eng GR - F31 NS056718/NS/NINDS NIH HHS/United States GR - K01 AG050719/AG/NIA NIH HHS/United States GR - T32 CA009547/CA/NCI NIH HHS/United States PT - Journal Article DEP - 20170218 PL - United States TA - JIMD Rep JT - JIMD reports JID - 101568557 PMC - PMC5680291 OTO - NOTNLM OT - Infantile Batten disease OT - Lysosomal storage disorders OT - Neuronal ceroid lipofuscinosis OT - Palmitoyl-protein thioesterase-1 OT - Soluble lysosomal enzymes EDAT- 2017/02/19 06:00 MHDA- 2017/02/19 06:01 PMCR- 2017/02/18 CRDT- 2017/02/19 06:00 PHST- 2016/09/09 00:00 [received] PHST- 2016/12/29 00:00 [accepted] PHST- 2016/12/13 00:00 [revised] PHST- 2017/02/19 06:00 [pubmed] PHST- 2017/02/19 06:01 [medline] PHST- 2017/02/19 06:00 [entrez] PHST- 2017/02/18 00:00 [pmc-release] AID - 1 [pii] AID - 10.1007/8904_2017_1 [doi] PST - ppublish SO - JIMD Rep. 2017;36:85-92. doi: 10.1007/8904_2017_1. Epub 2017 Feb 18.