Human iPSCs-Derived Endothelial Cells with Mutation in HNF1A as a Model of Maturity-Onset Diabetes of the Young

Abstract

Patients with HNF1A-maturity-onset diabetes of the young (MODY) often develop endothelial dysfunction and related microvascular complications, like retinopathy. As the clinical phenotype of HNF1A-MODY diabetes varies considerably, we used human induced pluripotent stem cells (hiPSCs) from two healthy individuals (control) to generate isogenic lines with mutation in HNF1A gene. Subsequently, control hiPSCs and their respective HNF1A clones were differentiated toward endothelial cells (hiPSC-ECs) and different markers/functions were compared. Human iPSC-ECs from all cell lines showed similar expression of CD31 and Tie-2. VE-cadherin expression was lower in HNF1A-mutated isogenic lines, but only in clones derived from one control hiPSCs. In the other isogenic set and cells derived from HNF1A-MODY patients, no difference in VE-cadherin expression was observed, suggesting the impact of the genetic background on this endothelial marker. All tested hiPSC-ECs showed an expected angiogenic response regardless of the mutation introduced. Isogenic hiPSC-ECs responded similarly to stimulation with pro-inflammatory cytokine TNF-α with the increase in ICAM-1 and permeability, however, HNF1A mutated hiPSC-ECs showed higher permeability in comparison to the control cells. Summarizing, both mono- and biallelic mutations of HNF1A in hiPSC-ECs lead to increased permeability in response to TNF-α in normal glycemic conditions, which may have relevance to HNF1A-MODY microvascular complications.

Integrated bioinformatic analysis of HNF1A in human cancers

“…These mutations may cause abnormal transcription and translation, eventually leading to changes in phenotype and function. 24 HNF1A was reported to interact with other genes and pathways in cancer, 25,26 which is consistent with our results. These phenomena may explain the distinct prognostic value of HNF1A in different types of cancers.…”

Hypoxia as a Driving Force of Pluripotent Stem Cell Reprogramming and Differentiation to Endothelial Cells

“…Importantly, these models provide human-relevant information on disease mechanisms or drug responses, as compared to various animal models. As discussed above, hiPSCs demonstrate the capacity to efficiently differentiate into various cell types including ECs [92]. Importantly, many studies have already confirmed that patient-specific hiPSC-derived ECs recapitulate the specific pathological phenotype observed in a particular disease, either with a known genetic background or complex etiology and systemic course [93,94].…”

“…GATA4 was identified as a potential target while surveying the genome of such patients, and its mutation was associated with impaired endothelial-to-mesenchymal transition, a process important for normal aortic valve formation [101]. Similarly, in our studies using hiPSCs isogenic lines generated through the introduction of mutation in hepatocyte nuclear factor 1A (HNF1A), we aimed to model the endothelial state of patients with maturity onset diabetes of the young (HNF1A-MODY) [92]. In our hands, hiPSC-ECs expressed typical endothelial markers, exerted angiogenic functionality, properly responded to inflammatory cytokines as well as shear stress (Figure 5).…”

Role of Heme-Oxygenase-1 in Biology of Cardiomyocytes Derived from Human Induced Pluripotent Stem Cells

“…hiPSC.1 expressed pluripotency markers: NANOG, OCT4, SSEA4, TRA-1-60, TRA-1-81 ( Supplementary Figure S2 A), and spontaneously differentiated in vitro via EBs into cells originating from three germ layers (mesoderm, endoderm, and ectoderm— Supplementary Figure S2B ). Karyotype of all hiPSCs was examined ( Supplementary Figure S3 ) Pluripotent properties of hiPSC.2 and hiPSC.3 were characterized in our previous studies [ 17 , 18 ] (respectively).…”

“…Verified hiPSCs were cultured as described previously [ 18 ]. hiPSCs were differentiated to cardiomyocytes utilising the GiWi protocol, described by Lian et al [ 8 ].…”