Fibrolamellar Carcinomas–Growth arrested by paracrine signals complexed with synthesized 3-O sulfated heparan sulfate oligosaccharides
Authors:
Wencheng Zhang a b c d Yongmei Xu e f, Xicheng Wang b c d, Tsunekazu Oikawa a ,Guowei Su f, Eliane Wauthier a, Guoxiu Wu b c d, Praveen Sethupathy g, Zhiying He b c d, Jian Liu e f, Lola M. Reid a h
Affiliation:
aDepartment of Cell Biology and Physiology, University of North Carolina, Chapel Hill, NC 27599, United States
bInstitute for Regenerative Medicine, Shanghai East Hospital, School of Life Sciences and Technology, Tongji University School of Medicine, Shanghai 200123, China
cShanghai Engineering Research Center of Stem Cells Translational Medicine, Shanghai 200335, China
dShanghai Institute of Stem Cell Research and Clinical Translation, Shanghai 200120, China
eDivision of Chemical Biology and Medicinal Chemistry, Eshelman School of Pharmacology, University of North Carolina, Chapel Hill, NC 27599, United States
fGlycan Therapeutics Corporation, 617 Hutton Street, Raleigh, NC 27606, United States
gDivision of Biomedical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, United States
hProgram in Molecular Biology and Biotechnology, School of Medicine, University of North Carolina, Chapel Hill, NC 27599, United States
Description:
Fibrolamellar carcinomas (FLCs), lethal tumors occurring in children to young adults, have genetic signatures implicating derivation from biliary tree stem cell (BTSC) subpopulations, co-hepato/pancreatic stem cells, involved in hepatic and pancreatic regeneration. FLCs and BTSCs express pluripotency genes, endodermal transcription factors, and stem cell surface, cytoplasmic and proliferation biomarkers.
The FLC-PDX model, FLC-TD-2010, is driven ex vivo to express pancreatic acinar traits, hypothesized responsible for this model's propensity for enzymatic degradation of cultures. A stable ex vivo model of FLC-TD-2010 was achieved using organoids in serum-free Kubota's Medium (KM) supplemented with 0.1% hyaluronans (KM/HA). Heparins (10 ng/ml) caused slow expansion of organoids with doubling times of ∼7–9 days. Spheroids, organoids depleted of mesenchymal cells, survived indefinitely in KM/HA in a state of growth arrest for more than 2 months. Expansion was restored with FLCs co-cultured with mesenchymal cell precursors in a ratio of 3:7, implicating paracrine signaling. Signals identified included FGFs, VEGFs, EGFs, Wnts, and others, produced by associated stellate and endothelial cell precursors.
Fifty-three, unique heparan sulfate (HS) oligosaccharides were synthesized, assessed for formation of high affinity complexes with paracrine signals, and each complex screened for biological activity(ies) on organoids. Ten distinct HS-oligosaccharides, all 10–12 mers or larger, and in specific paracrine signal complexes elicited particular biological responses. Of note, complexes of paracrine signals and 3-O sulfated HS-oligosaccharides elicited slowed growth, and with Wnt3a, elicited growth arrest of organoids for months. If future efforts are used to prepare HS-oligosaccharides resistant to breakdown in vivo, then [paracrine signal—HS-oligosaccharide] complexes are potential therapeutic agents for clinical treatments of FLCs, an exciting prospect for a deadly disease.