Control Stem Cell Research Articles

Identification of Domains and Factors Involved in MINIYO Nuclear Import.

The transition of stem cells from self-renewal into differentiation is tightly regulated to assure proper development of the organism. Arabidopsis MINIYO (IYO) and its mammalian orthologue RNA polymerase II associated protein 1 (RPAP1) are essential factors for initiating stem cell differentiation in plants and animals. Moreover, there is evidence suggesting that the translocation of IYO and RPAP1 from the cytosol into the nucleus functions as a molecular switch to initiate this cell fate transition. Identifying the determinants of IYO subcellular localization would allow testing if, indeed, nuclear IYO migration triggers cell differentiation and could provide tools to control this crucial developmental transition. Through transient and stable expression assays in Nicotiana benthamiana and Arabidopsis thaliana, we demonstrate that IYO contains two nuclear localization signals (NLSs), located at the N- and C-terminus of the protein, which mediate the interaction with the NLS-receptor IMPA4 and the import of the protein into the nucleus. Interestingly, IYO also interacts with GPN GTPases, which are involved in selective nuclear import of RNA polymerase II. This interaction is prevented when the G1 motif in GPN1 is mutated, suggesting that IYO binds specifically to the nucleotide-bound form of GPN1. In contrast, deleting the NLSs in IYO does not prevent the interaction with GPN1, but it interferes with import of GPN1 into the nucleus, indicating that IYO and GPN1 are co-transported as a complex that requires the IYO NLSs for import. This work unveils key domains and factors involved in IYO nuclear import, which may prove instrumental to determine how IYO and RPAP1 control stem cell differentiation.

Effects of exercise training and stem cell therapy on the left ventricle of infarcted rats

Introduction and ObjectivesStem cell therapy and aerobic exercise are non-pharmacological therapies following myocardial infarction. The aim of this study was to test whether aerobic exercise training enhances the benefits of mesenchymal stem cell (MSC) therapy on remodeling of the extracellular matrix and fetal gene expression in the left ventricle of infarcted rats. MethodsMyocardial infarction was surgically induced in six-week old male Wistar rats. Animals were divided into four groups: sedentary control (SC) and sedentary and stem cell treated (SCMSC); exercised (EX) and exercised and stem cell treated (EXMSC). Bone marrow-derived MSCs were immediately transplanted via the tail vein (concentration: 1×106 cells). Exercise training (five days/week, 60 min/day; 60% of maximal running speed) started 24 hours after myocardial infarction and lasted for 12 weeks. ResultsExercise capacity was higher in exercised than in sedentary groups. Animals in the SCMSC, EX and EXMSC groups exhibited better cardiac function than those in SC. Collagen content was lower in the SCMSC, EX and EXMSC groups than in SC and skeletal α-actin expression was lower in EX and EXMSC than in SC. The α/β-MHC ratio was higher in EX and EXMSC than in SC. The combination of therapies further reduced collagen content in the remote region of the infarct (∼24%) and skeletal α-actin expression (∼30%). ConclusionAerobic exercise training appears to enhance the beneficial effects of stem cell therapy on remodeling of the extracellular matrix and fetal gene expression in the left ventricle of rats with moderate infarction.

Orchestrating stem cell fate: Novel tools for regenerative medicine

Mesenchymal stem cells are undifferentiated cells able to acquire different phenotypes under specific stimuli. In vitro manipulation of these cells is focused on understanding stem cell behavior, proliferation and pluripotency. Latest advances in the field of stem cells concern epigenetics and its role in maintaining self-renewal and differentiation capabilities. Chemical and physical stimuli can modulate cell commitment, acting on gene expression of Oct-4, Sox-2 and Nanog, the main stemness markers, and tissue-lineage specific genes. This activation or repression is related to the activity of chromatin-remodeling factors and epigenetic regulators, new targets of many cell therapies. The aim of this review is to afford a view of the current state of in vitro and in vivo stem cell applications, highlighting the strategies used to influence stem cell commitment for current and future cell therapies. Identifying the molecular mechanisms controlling stem cell fate could open up novel strategies for tissue repairing processes and other clinical applications.

Unfolded protein response-mediated modulation of mesenchymal stem cells.

The endoplasmic reticulum (ER) receives unfolded proteins predestined for the secretory pathway or to be incorporated as transmembrane proteins. The ER has to accommodate the proper folding and glycosylation of these proteins and also to properly incorporate transmembrane proteins. However, under various circumstances, the proteins shuttling through the ER can be misfolded and undergo aggregation, which causes activation of the unfolded protein response (UPR). The UPR is mediated through three primary pathways: activating transcription factor-6, inositol-requiring enzyme-1 (IRE1), and PKR-like endoplasmic reticulum kinase, which up-regulate ER folding chaperones and temporarily suppress protein translation. The UPR can be both cytoprotective and/or cytotoxic depending on the duration of UPR activation and the type of host cell. Proteostasis controls stem cell function, while stress responses affect stem cell identity and differentiation. The present review aimed to explore and discuss the effects of the UPR pathways on mesenchymal stem cells.

The RNA hairpin binder TRIM71 modulates alternative splicing by repressing MBNL1.

TRIM71/LIN-41, a phylogenetically conserved regulator of development, controls stem cell fates. Mammalian TRIM71 exhibits both RNA-binding and protein ubiquitylation activities, but the functional contribution of either activity and relevant primary targets remain poorly understood. Here, we demonstrate that TRIM71 shapes the transcriptome of mouse embryonic stem cells (mESCs) predominantly through its RNA-binding activity. We reveal that TRIM71 binds targets through 3' untranslated region (UTR) hairpin motifs and that it acts predominantly by target degradation. TRIM71 mutations implicated in etiogenesis of human congenital hydrocephalus impair target silencing. We identify a set of primary targets consistently regulated in various human and mouse cell lines, including MBNL1 (Muscleblind-like protein 1). MBNL1 promotes cell differentiation through regulation of alternative splicing, and we demonstrate that TRIM71 promotes embryonic splicing patterns through MBNL1 repression. Hence, repression of MBNL1-dependent alternative splicing may contribute to TRIM71's function in regulating stem cell fates.

Human Carcinogenesis: The Role of Age and Gender

To identify the reason for age and gender differences in cancer risk. Age-standardized incidence rates for 17 cancer types were compared between genders in 50 populations. For each cancer type, the female/male rate ratio was listed in fixed order of population. Correlation coefficients were calculated between these lists in all pairwise combinations. For each population, the female/male rate ratio was listed in fixed order of cancer. Correlation coefficients were calculated between lists in all pairwise combinations. Only four pairwise combinations for cancer type gave a correlation coefficient greater than 0.700. For each population, the lowest correlation coefficient was 0.950. The reason for the differences in risk of cancer varies with each type of cancer, but remains fixed in all populations. It is suspected that species-specific genes control stem cell telomere dynamics in a fixed strategy at rates that vary among tissues and between genders.

Nondestructive Characterization of Stem Cell Neurogenesis by a Magneto-Plasmonic Nanomaterial-Based Exosomal miRNA Detection.

The full realization of stem cell-based treatments for neurodegenerative diseases requires precise control and characterization of stem cell fate. Herein, we report a multifunctional magneto-plasmonic nanorod (NR)-based detection platform to address the limitations associated with the current destructive characterization methods of stem cell neurogenesis. Exosomes and their inner contents have been discovered to play critical roles in cell-cell interactions and intrinsic cellular regulations and have received wide attention as next-generation biomarkers. Moreover, exosomal microRNAs (miRNA) also offer an essential avenue for nondestructive molecular analyses of cell cytoplasm components. To this end, our developed nondestructive, selective, and sensitive detection platform has (i) an immunomagnetic active component for exosome isolation and (ii) a plasmonic/metal-enhanced fluorescence component for sensitive exosomal miRNA detection to characterize stem cell differentiation. In a proof-of-concept demonstration, our multifunctional magneto-plasmonic NR successfully detected the expression level of miRNA-124 and characterized neurogenesis of human-induced pluripotent stem cell-derived neural stem cells in a nondestructive and efficient manner. Furthermore, we demonstrated the versatility and feasibility of our multifunctional magneto-plasmonic NRs by characterizing a heterogeneous population of neural cells in an ex vivo rodent model. Collectively, we believe our multifunctional magneto-plasmonic NR-based exosomal miRNA detection platform has a great potential to investigate the function of cell-cell interactions and intrinsic cellular regulators for controlling stem cell differentiation.

Tunable Chemical and Topographic Patterns Based on Binary Colloidal Crystals (BCCs) to Modulate MG63 Cell Growth

AbstractMore effective and diversified surface modification strategies are required for materials used in biomedical engineering. Combining surface modification involving bioactive signals or nonfouling polymers with tunable topography has the potential to meet this need. Here, a method is reported to generate bioactive surfaces having tunable topographies based on self‐assembled binary colloidal crystals (BCCs), where the colloids are premodified with nonfouling molecules or cell adhesive peptides. The BCCs are fabricated from silica (Si) microspheres and polymer nanospheres. The Si microspheres are either modified with poly(ethylene glycol) (PEG) or with the cell adhesive arginine–glycine–aspartic acid (RGD) peptide prior to BCC fabrication. Four types of BCCs are explored in cell studies using MG63 cells. BCC surfaces coupled with PEG or RGD peptides are found to significantly modulate cell adhesion, spreading, and morphology, which is attributed to the combination of BCC topography and the molecules at the particle surface within the BCC. PEG‐modified BCCs are expected to find applications where limited cell adhesion is required, while the RGD‐modified BCCs have the potential for enhancing cell growth on medical devices such as bone implants. More advanced cell biology applications such as controlled stem cell differentiation are also anticipated to find use from BCCs.

Targeting cancer stem cells in drug discovery: Current state and future perspectives.

In recent decades, cancer stem cells (CSCs) have been increasingly identified in many malignancies. CSC-related signaling pathways and their functions provide new strategies for treating cancer. The aberrant activation of related signaling pathways (e.g., Wnt, Notch, and Hedgehog pathways) has been linked to multiple types of malignant tumors, which makes these pathways attractive targets for cancer therapy. CSCs display many characteristic features, such as self-renewal, differentiation, high tumorigenicity, and drug resistance. Therefore, there is an urgent need to develop new therapeutic strategies to target these pathways to control stem cell replication, survival, and differentiation. Notable crosstalk occurs among different signaling pathways and potentially leads to compensatory escape. Therefore, multitarget inhibitors will be one of the main methods to overcome the drug resistance of CSCs. Many small molecule inhibitors of components of signaling pathways in CSCs have entered clinical trials, and some inhibitors, such as vismodegib, sonidegib, and glasdegib, have been approved. Tumor cells are susceptible to sonidegib and vismodegib resistance due to mutations in the Smo protein. The signal transducers and activators of transcription 3 (STAT3) inhibitor BBI608 is being evaluated in a phase III trial for a variety of cancers. Structural derivatives of BBI608 are the main focus of STAT3 inhibitor development, which is another strategy for CSC therapy. In addition to the potential pharmacological inhibitors targeting CSC-related signaling pathways, other methods of targeting CSCs are available, such as nano-drug delivery systems, mitochondrion targeting, autophagy, hyperthermia, immunotherapy, and CSC microenvironment targeting. In addition, we summarize the latest advances in the clinical development of agents targeting CSC-related signaling pathways and other methods of targeting CSCs.

Fabrication of High-resolution Graphene-based Flexible Electronics via Polymer Casting

In this study, a novel method based on the transfer of graphene patterns from a rigid or flexible substrate onto a polymeric film surface via solvent casting was developed. The method involves the creation of predetermined graphene patterns on the substrate, casting a polymer solution, and directly transferring the graphene patterns from the substrate to the surface of the target polymer film via a peeling-off method. The feature sizes of the graphene patterns on the final film can vary from a few micrometers (as low as 5 µm) to few millimeters range. This process, applied at room temperature, eliminates the need for harsh post-processing techniques and enables creation of conductive graphene circuits (sheet resistance: ~0.2 kΩ/sq) with high stability (stable after 100 bending and 24 h washing cycles) on various polymeric flexible substrates. Moreover, this approach allows precise control of the substrate properties such as composition, biodegradability, 3D microstructure, pore size, porosity and mechanical properties using different film formation techniques. This approach can also be used to fabricate flexible biointerfaces to control stem cell behavior, such as differentiation and alignment. Overall, this promising approach provides a facile and low-cost method for the fabrication of flexible and stretchable electronic circuits.

Ca2+ Signaling and Regeneration.

Regeneration is the process by which lost or damaged tissue is replaced in adult organisms. Some organisms exhibit robust regenerative capabilities, while others, including humans, do not. Understanding the molecular principles governing the regenerative malleability of different organisms is of fundamental biological interest. Further, this problem has clear impact for the field of "regenerative medicine," which aspires to understand how human cells, tissues, and organs may be restored to normal function in scenarios of disease, damage, or age-related decline. This review will focus on the planarian flatworm as a powerful model system for studying the role of Ca2+ signals in regeneration. These invertebrate animals display an astounding innate regenerative capacity capable of regenerating complete organisms from tiny, excised fragments. New knowledge and methodological capabilities in this system highlight the potential for studying the role of Ca2+ signaling at multiple stages of the regenerative blueprint that controls stem cell behavior in vivo.

Putting Stem Cells on a Low-Fat Diet Switches Their Pluripotent State.

Previous work demonstrating the importance of stem cell metabolism in regulating development ignited a debate on how intrinsic versus extrinsic conditions influence metabolic programming of epigenetics. In this issue of Cell Stem Cell, Cornacchia etal. show that exogenous lipids regulate an acetyl-CoA-dependent switch to influence the intrinsic pluripotent state of hPSCs.

In vivo administration of G9a inhibitor A366 decreases osteogenic potential of bone marrow-derived mesenchymal stem cells.

Epigenetic mechanisms such as histone methylation are considered as one of the most important mediators that control stem cell behaviors such as proliferation, senescence and differentiation. G9a, a histone methyltransferase, has recently generated intense attention as potential target for controlling many diseases such as cancers. The aim of the present study was to evaluate the effect of in vivo administration of A366, a G9a inhibitor, on proliferative and differentiation potential of bone marrow-derived mesenchymal stem cells (BM-MSCs). We inhibited G9a using intraperitoneally administration of A366, and we evaluated BM-MSC proliferation and differentiation behaviors in vitro. Colony formation assay of BM-MSCs at primary culture showed that in vivo administration of A366 reduced the colony forming capacity of BM-MSCs. Moreover, PDT of BM-MSC isolated from A366-treated rats was higher than control, especially in the early passages. BM-MSC isolated from A366-treated rats showed higher adipogenic potential compared to the control at the early passages as determined by gene expression and Oil Red staining. Whereas, osteogenic potential of BM-MSC isolated from A366-treated rats was lower than control, especially at early passages. Our results suggest that the epigenetic modifier such as A366, which seems to be a therapeutic approach for controlling diseases such as cancer, might also influence the proliferation and differentiation capacity of MSCs both in vitro and in vivo. Moreover, epigenetic modifying chemicals seem to be a strategy to manipulate MSC expansion capacity and differentiation propensity, as well as to efficiently involvement of MSCs in tissue homeostasis, cell-based therapy and tissue engineering.

Cord blood research, banking, and transplantation: achievements, challenges, and perspectives.

The first hematopoietic transplant in which umbilical cord blood (UCB) was used as the source of hematopoietic cells was performed in October 1988. Since then, significant achievements have been reported in terms of our understanding of the biology of UCB-derived hematopoietic stem (HSCs) and progenitor (HPCs) cells. Over 40,000 UCB transplants (UCBTs) have been performed, in both children and adults, for the treatment of many different diseases, including hematologic, metabolic, immunologic, neoplastic, and neurologic disorders. In addition, cord blood banking has been developed to the point that around 800,000 units are being stored in public banks and more than 4 million units in private banks worldwide. During these 30 years, research in the UCB field has transformed the hematopoietic transplantation arena. Today, scientific and clinical teams are still working on different ways to improve and expand the use of UCB cells. A major effort has been focused on enhancing engraftment to potentially reduce risk of infection and cost. To that end, we have to understand in detail the molecular mechanisms controlling stem cell self-renewal that may lead to the development of ex vivo systems for HSCs expansion, characterize the mechanisms regulating the homing of HSCs and HPCs, and determine the relative place of UCBTs, as compared to other sources. These challenges will be met by encouraging innovative research on the basic biology of HSCs and HPCs, developing novel clinical trials, and improving UCB banking both in the public and private arenas.

Efficacies of stem cell therapies for functional improvement of the β cell in patients with diabetes: A systematic review of controlled clinical trials.

Background & Aim This study was performed to investigate whether stem cell therapy enhances β cell function by meta-analysis with proper consideration of variability of outcome measurements in controlled trial of type 1 diabetes mellitus (T1DM) and type 2 diabetes mellitus (T2DM) patients. Methods, Results & Conclusion Method A systematic search was performed from inception to January 2018 in PubMed, EMBASE, and Cochrane databases. Β cell function was assessed by stimulated C-peptide, fasting C-peptide, normal glycosylated hemoglobin levels (HbA1C), and exogenous insulin dose patterns. The quality of the studies were assessed by both the Cochrane Collaboration's Risk of Bias (ROB) for Randomized controlled trials and the Risk of Bias in Non-randomized Studies of Interventions (ROBINS-I) for non-randomized controlled trials. Result From the selected final 15 articles, total of 16 trials were analyzed. There were 6 T1DM trials (total 153 cases) and 10 T2DM trials (total 457 cases). In T2DM patients, the changes in stimulated C-peptide, HbA1c, and exogenous insulin dose versus baseline showed a favorable pattern with significant heterogeneity in stem cell therapy. In T1DM, there was no significant difference between control group and stem cell therapy group in three indicators except for HbA1c. Most of the studies were rated as having high risk of bias in the quality assessment. Conclusion The stem cell therapy for DM patients is not effective in T1 DM but seems to be effective in improving the β cell function in T2 DM. Further verification through a rigorously designed study is warranted.

Towards cell therapy of polycystic ovary syndrome (PCOS): human mesenchymal stem cells secretome inhibits androgen production by pcos theca cells

Background & Aim Background Polycystic Ovarian Syndrome (PCOS) is a metabolic disorder characterized by inflammation, infertility and excess ovarian androgen production by theca cells. Though its etiology is not fully understood. Women with PCOS exhibit increased expression of CYP17A1, and CYP11A1 genes involved in androgen production. Due to the robust anti-inflammatory ability of human bone marrow Mesenchymal stem cells (hMSCs), we evaluated its utility as a novel cell-therapy by injecting hMSCs directly into the ovaries of PCOS mouse model. It showed a significant improvement in hyperandrogenemia. hMSCs also secrete various factors in media, known as secretome. The literature shows that the secretome of hMSCs has the same regenerative and reparative function likes hMSCs. Hypothesis We hypothesize that hMSCs secretome contains the factors that are able to inhibit the androgen biosynthesis in PCOS theca cells. Methods, Results & Conclusion Material and Methods Theca cells, collected from normal cycling and PCOS patient underwent ovariectomy, were seeded on culture dishes pre-coated with the extracellular matrix at a density of 6 × 104 cells/ml and cultured for 24 hours. Cells were then treated with either 3T3 conditioned media as a control (C) or Mesenchymal stem cells conditioned media (CM). The C and CM were prepared through collecting the serum-free supernatant of 3T3 and hMSCs at an optimal confluence, 48 hours later. The mRNA and protein expression of CYP17A1, CYP11A1, and DENND1A.V2 were quantified by RT-PCR and Western blot, while testosterone level in media was measured by RIA assay. Results Human PCOS theca cells treated with hMSCs CM for 48 hours secreted significantly lower level of testosterone (146 ± 13.6 ng/dl) compared to control C (215 ± 11.3 ng/dl) (P Conclusion The therapeutic capabilities of hMSCs in PCOS are partially mediated by its secretome which contains various growth factors and anti-inflammatory cytokines both free and within extracellular vesicles. Stem cell therapy, either by cell implantation or via secretome injection might potentially be a novel approach for effective treatment of PCOS patients.

Noncanonical NF-κB controls stem cell signatures in the colonic mucosa and affects susceptibility to inflammation-induced carcinogenesis

Abstract NF-κB signaling is divided into two distinct pathways, defined as either canonical or noncanonical. Canonical NF-κB signaling is well described and characterized. However, significantly less is known regarding mechanisms regulated by the noncanonical NF-κB pathway, especially outside of the lymphoid system. Recently, interest in the role of noncanonical NF-κB in the mucosal immune system has been increasing, with emphasis on NF-κB inducing kinase (NIK). NIK is an essential kinase in this cascade. Our data shows that lack of noncanonical signaling through the complete knockout of Nik in mice results in an aberrant large intestine phenotype characterized by reduced stem cell marker expression, altered regeneration and differentiation capacity under stress, changes in the microbiome, and predisposition to inflammation-associated tumorigenesis. Subsequent mechanistic studies utilizing novel mice with cell type specific deletions of either RelA/p65 or Nik revealed separate and distinct rolls for canonical and noncanonical NF-κB signaling in attenuating inflammation and tumor progression. RelA/p65 attenuates disease progression through the myeloid compartment in the gut, whereas Nik functions through the intestinal epithelial cell compartment. Dysregulation of noncanonical signaling is also observed in human colorectal cancer patients, which have marked suppression of this pathway in colonic biopsy samples. Together, these findings indicate that noncanonical NF-κB signaling is a critical regulator of immune system homeostasis in the gastrointestinal tract and plays a vital protective role against colorectal cancer.

Efficacies of Stem Cell Therapies for Functional Improvement of the β Cell in Patients with Diabetes: A Systematic Review of Controlled Clinical Trials.

Background and ObjectivesThis study was performed to investigate whether stem cell therapy enhances β cell function by meta-analysis with proper consideration of variability of outcome measurements in controlled trial of type 1 diabetes mellitus (T1DM) and type 2 diabetes mellitus (T2DM) patients.MethodsA systematic search was performed from inception to January 2018 in PubMed, EMBASE, and Cochrane databases. β cell function was assessed by stimulated C-peptide, fasting C-peptide, normal glycosylated hemoglobin levels (HbA1C), and exogenous insulin dose patterns. The quality of the studies were assessed by both the Cochrane Collaboration’s Risk of Bias (ROB) for Randomized controlled trials and the Risk of Bias in Non-randomized Studies of Interventions (ROBINS-I) for non-randomized controlled trials.ResultsFrom the selected final 15 articles, total of 16 trials were analyzed. There were 6 T1DM trials (total 153 cases) and 10 T2DM trials (total 457 cases). In T2DM patients, the changes in stimulated C-peptide, HbA1c, and exogenous insulin dose versus baseline showed a favorable pattern with a significant heterogeneity in stem cell therapy. In T1DM, there was no significant difference between control group and stem cell therapy group in three indicators except for HbA1c. Most of the studies were rated as having high risk of bias in the quality assessment.ConclusionsThe stem cell therapy for DM patients is not effective in T1DM but seems to be effective in improving the β cell function in T2DM. However the observed effect should be interpreted with caution due to the significant heterogeneity and high risk of bias within the studies. Further verification through a rigorously designed study is warranted.

Spatially Restricted Stromal Wnt Signaling Restrains Prostate Epithelial Progenitor Growth through Direct and Indirect Mechanisms.

Cell-autonomous Wnt signaling has well-characterized functions in controlling stem cell activity,including in the prostate. While niche cells secreteWnt ligands, the effects of Wnt signaling in niche cells per se are less understood. Here,we show that stromal cells in the proximal prostatic duct near the urethra, a mouse prostate stem cell niche, not only produce multiple Wnt ligands but also exhibit strong Wnt/β-catenin activity. The non-canonical Wnt ligand Wnt5a, secreted by proximal stromal cells, directly inhibits proliefration of prostateepithelial stem or progenitor cells whereas stromal cell-autonomous canonical Wnt/β-catenin signaling indirectly suppresses prostate stem or progenitor activity via the transforming growth factor β (TGFβ) pathway. Collectively, these pathways restrain the proliferative potential of epithelial cells in the proximal prostatic ducts. Human prostate likewise exhibits spatially restricted distribution of stromal Wnt/β-catenin activity, suggesting a conserved mechanism for tissue patterning. Thus, this study showshow distinct stromal signaling mechanisms within theprostate cooperate to regulate tissue homeostasis.

O1.5. SCHIZOPHRENIA PATIENT-DERIVED STEM CELLS FOR SYSTEMS GENOMICS: GENETIC VARIANTS, MOLECULAR PATHWAYS AND CELL FUNCTIONS ALTERED IN INDIVIDUAL PATIENTS

Background: Genome-wide association studies have identified over 100 loci in which common genetic variants are associated with schizophrenia in the population. Other rare genetic risk variants have also been identified. The challenge to the field is to understand the functional consequences of these genetic variants and the frequency and combinations of common and rare variants in individuals are unknown. How do we move to a functional genomics understanding of schizophrenia? This must come from the biology of individual patient-derived cells where genotype relates directly to cell phenotype and where specific genetic variants are examined in context of the individual’s full array of genetic risks. Methods: We have developed a novel systems-based approach using multi-omics data and cellular function assays using schizophrenia patient-derived olfactory neural stem cells as the model system. We are identifying genetic variants that influence schizophrenia-associated cell functions in individual patient cells. This integrative and deep analysis of molecular networks, signaling pathways and cell functions in the same individual patient and control stem cells is unique in the fields of schizophrenia, cell biology, and systems biology. Results: Whole-exome sequencing data from this cohort of patient-derived stem cells confirm that the patient cells have higher polygenic risk scores when compared to healthy controls. We filtered schizophrenia-relevant, rare and common variants, present in individual patients, by their participation in the schizophrenia protein-protein interaction network that we previously built using genes located in schizophrenia-associated loci. We found that these selected variants were significantly enriched in schizophrenia-associated gene networks, pathways and functions that we have previously identified in these patient-derived cells: i.e. in multiple regulatory layers including DNA methylation, transcriptome and proteome; and in multiple assays of cell functions including cell proliferation, cell cycle, cell adhesion, cell motility, and protein synthesis. Discussion: This systems-level view draws attention away from single candidate genes and single cell functions, identifying failure of regulation of networks and pathways as central to schizophrenia stem cell biology, leading to modulation of multiple cell functions. Such failures of regulation could change subtly the behaviour of cells and the assembly of the brain during development as well as cell functions in the mature brain. Our long-term goal is to use un-biased systems-level cellular and molecular data from patient-derived stem cells to build a roadmap for discovery of new therapeutic strategies tailored to individual’s molecular and cellular signatures.