title: A simple, reliable method for high-throughput screening for diabetes drugs using 3D β-cell spheroids.
abstract: Early screens for new diabetes drugs rely on monolayers of β-cells, which are known to be poor predictors of the in vivo response. Previously, we developed a method to create uniform islet spheroids from freshly-dispersed human donor tissue for drug screening. While the human engineered islets worked well to reduce donor-to-donor variability, it is difficult and expensive to obtain sufficient high-quality human islets for drug testing. Thus, this study utilized a genetically-modified β-cell culture line (INS-1832/13) in 2D and as 3D spheroids and compared the results to human islet tissue formed into spheroids using a high-throughput 384-well format. In response to increasing concentrations of glucose, all 3 groups increased insulin release, but the cultured β-cells (2D and 3D) were more sensitive to glucose (EC50 5.85mM for 2D β-cells, 16.24mM for 3D β-cell spheroids) than the human islet spheroids (EC50 53.69mM). The order of responses to glybenclamide was human spheroids >3D β-cell culture >2D β-cell culture. In response to caffeine, the β-cells in 2D or 3D were more responsive compared to the human islet spheroids (EC50 0.39 and 0.31mM for 2D and 3D β-cells respectively). When exposed to inhibitors of insulin secretion (nifedipine and diazoxide), the responses were more similar between groups. Z' calculations, indicative of assay quality, determined that the 3D β-cell spheroids reached the criteria of an excellent to ideal drug screen assay more consistently than the other test models. In conclusion, 3D β-cell spheroids from a cultured cell line can be used in HTS assays that, according to reference drugs tested here, are sensitive and predictive of the in vivo response.
authors.affiliation: University of Missouri-Kansas City School of Medicine, 2464 Charlotte St., Kansas City, MO 64108, United States.
authors.forename: Jesal
authors.initials: J
authors.lastname: Amin
authors.affiliation: Likarda, LLC, 2002 W 39th Ave, Kansas City, KS 66103, United States.
authors.forename: Karthik
authors.initials: K
authors.lastname: Ramachandran
authors.affiliation: University of Kansas Medical Center, Department of Physical Therapy and Rehabilitation Science, 3901 Rainbow Blvd., Kansas City, KS 66160, United States; Likarda, LLC, 2002 W 39th Ave, Kansas City, KS 66103, United States.
authors.forename: S Janette
authors.initials: SJ
authors.lastname: Williams
authors.affiliation: Rockhurst University, Department of Chemistry, 1100 Rockhurst Rd, Kansas City, MO 64110, United States.
authors.forename: Annie
authors.initials: A
authors.lastname: Lee
authors.affiliation: University of Kansas Medical Center, Department of Physical Therapy and Rehabilitation Science, 3901 Rainbow Blvd., Kansas City, KS 66160, United States.
authors.forename: Lesya
authors.initials: L
authors.lastname: Novikova
authors.affiliation: University of Kansas Medical Center, Department of Physical Therapy and Rehabilitation Science, 3901 Rainbow Blvd., Kansas City, KS 66160, United States; Likarda, LLC, 2002 W 39th Ave, Kansas City, KS 66103, United States. Electronic address: lbittel@kumc.edu.
authors.forename: Lisa
authors.initials: L
authors.lastname: Stehno-Bittel
mesh_terms: D000328:Adult; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D055808:Drug Discovery / Q000379:methods*; D004353:Drug Evaluation, Preclinical; D005947:Glucose / Q000494:pharmacology; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007004:Hypoglycemic Agents / Q000737:chemistry / Q000494:pharmacology*; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000187:drug effects* / Q000378:metabolism; D018874:Spheroids, Cellular / Q000187:drug effects* / Q000378:metabolism
chemical_list: D007004:Hypoglycemic Agents; D007328:Insulin; D005947:Glucose
keywords: Caffeine; Diabetes; Diazoxide; Glucose; Glybenclamide; High-throughput screen; Nifedipine; β-cell