Exploring mood disorders and treatment options using human stem cells

Hudock, Autumn; Leal, Zaira Paulina; Sharma, Amandeep; Mei, Arianna; Santos, Renata; Marchetto, Maria Carolina

doi:10.1590/1678-4685-GMB-2023-0305

person Autumn Hudock

conceptualized the manuscript · supervised the other authors · performed literature research · curated the bibliography · wrote the manuscript · designed and generated figures and tables · reviewed and edited the manuscript · approved the final version of the manuscript

schoolUniversity of California San Diego, Department of Anthropology, La Jolla, CA, USA.

0000-0002-4722-754X

person Zaira Paulina Leal

performed literature research · curated the bibliography · wrote the manuscript · designed and generated figures and tables · reviewed and edited the manuscript · approved the final version of the manuscript

schoolUniversity of California San Diego, Department of Anthropology, La Jolla, CA, USA.

0009-0002-2902-9248

person Amandeep Sharma

designed and generated figures and tables · analyzed the data · wrote the methodology session · reviewed and edited the manuscript · approved the final version of the manuscript

schoolThe Salk Institute for Biological Studies, Laboratory of Genetics, La Jolla, CA, USA.

0000-0002-2091-9618

person Arianna Mei

conducted the experiments · analyzed the data · reviewed and edited the manuscript · approved the final version of the manuscript

schoolThe Salk Institute for Biological Studies, Laboratory of Genetics, La Jolla, CA, USA.

person Renata Santos

conducted the experiments · reviewed and edited the manuscript · approved the final version of the manuscript

schoolThe Salk Institute for Biological Studies, Laboratory of Genetics, La Jolla, CA, USA.schoolUniversité Paris Cité, Institute of Psychiatry and Neuroscience of Paris (IPNP), INSERM U1266, Signaling Mechanisms in Neurological Disorders, Paris, France.schoolInstitut des Sciences Biologiques, Centre National de la Recherche Scientifique (CNRS), Paris, France.

0000-0002-3085-5128

person Maria Carolina Marchetto

conceptualized the manuscript · supervised the other authors · approved the final version of the manuscript

schoolUniversity of California San Diego, Department of Anthropology, La Jolla, CA, USA.

0000-0002-0449-9051

Send correspondence to Maria Carolina Marchetto. University of California San Diego, Department of Anthropology, 9500 Gilman Drive, MC 0532, La Jolla, CA, USA. E-mail:
mcmarchetto@ucsd.edu.

Associate Editor:

Alberto R. Kornblihtt

Conflict of Interest

The authors declare that there is no conflict of interest that could be perceived as prejudicial to the impartiality of the reported research.

Figures (3)
Tables (2)

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Figure 1 - Using patient-iPSC models to study mood disorders. Patient skin or blood cells are collected and reprogrammed into iPSC using previously established protocols (see methods). The iPSCs are then differentiated into neuronal or glial progenitor cells in 2 or 3D cultures and matured into functional neurons and glial cells. These cells can then be used to study the disease pathophysiology, response to inflammatory stimuli and drug screening for novel therapeutic compounds. The graph represents the protocol of establishing cell lines in tissue culture settings to study mood disorders affecting the brain, beginning with the recruitment of cells, the process of reprogramming cells into iPSCs, and differentiating them into various neuronal cell types.

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Figure 2 - Apigenin attenuated the inflammation response more effectively in iPSCs-derived astrocytes from BD patients. (A) Schematics of investigating anti-inflammatory attributes of a flavonoid (Apigenin). iPSC, glial progenitor cells (GPCs) and astrocytes derived from three neurotypical and six BD donors, described previously (Santos et al., 2017; Vadodaria et al., 2021) were used in this study. This assay was previously described by Vadodaria et al. (2021). Briefly, pro-inflammatory stimuli were added to 4-week-old astrocytes for 5 hours, either by using recombinant human IL-1b (10 ng/mL) or recombinant human TNF-a (50 ng/mL); PBS was used as a non-stimulated control. To mitigate the inflammatory response, astrocytes were simultaneously treated with 20 μM Apigenin. BD GolgiPlug and BD GolgiStop were added to the treatments to inhibit extracellular protein secretion. Flow cytometry was used to quantify IL-6 producing cells. Astrocytes were dissociated using a 1:1 ratio of accutase/papain followed by staining with Zombie UV fixable Viability kit. The BD Cytofix/Cytoperm and BD Perm/Wash kits were used for fixing and permeabilizing the cells. Subsequently, IL-6 cytokine was labeled with APC conjugated anti-IL-6 antibody in BD Perm/Wash for 20 min. Data collection and analysis were conducted using a BD CantoII cytometer and FlowJo software, respectively. Negative gating controls for anti-IL-6 were done in non-stimulated samples stained with rat IgG1-APC antibody. The IL-6 positive cells were quantified by normalizing the data with non-stimulated cells from the vehicle treated cells. Data are from two biological experiments with technical replicates (3). (B & C) Bar graph representing the quantification of astrocytes expressing IL-6 cytokine post 5 hours of exposure to Vehicle or IL1-b or IL1-b+Apigenin (B) or Vehicle or TNF-a or TNF-a+Apigenin (C), mean ± SEM. Two-way Anova test was used to determine the statistical significance.

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Figure 3 - Exploring alternative treatment methods for BD and MDD non-responders to lithium and SSRIs/ SNRIs using iPSC models. The graph depicts the pathways that researchers have used to explore alternative treatment methods beyond lithium and SSRIs/SNRIs for non-responders with BD and MDD, as they make up about half of the population for those diagnosed with both disorders (Tighe et al., 2011; Taliaz et al., 2021). Beginning with the collection of cells, researchers can look for biomarkers associated with drug responsiveness, and model personalized neuronal models of patients that can be subjected to various forms of analysis. This data can contribute to the exploration of new compounds and alternative drugs already on the market, leading to relief for non-responders of traditional treatment methods.

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Table 1 - Induced pluripotent stem cells in the study of bipolar disorder: a chronological table of exploratory and treatment-based studies.

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Table 2 - Induced pluripotent stem cells in the study of major depressive disorder: a chronological table of exploratory and treatment based studies.

Study Type	Treatment	Cell/ Tissue Type(s)	iPSCs Relevance	Findings	Reference
Exploratory	N/a	iPSCs Neurons	Investigation of the cellular behavior and developmental pathways of iPSC derived neurons from BD patients and controls	-Increased membrane receptors and ion-controlled genes were noted in BD patient derived neurons compared to controls	Chen et al. (2014)
Exploratory	N/a	Postmortem cerebral tissue iPSCs Neurons NPCs	Examination of the expression of hsa-miR-34a in postmortem tissue and various IPSCs derived cell types	-Increased expression of microRNAs in BD patient derived iPSC cultures and postmortem cerebral tissue	Bavamian et al. (2015)
Exploratory	N/a	iPSCs NPCs Neurons	Models of iPSCs, NPCs and neurons of a 1^st degree family cohort analyzed using microarray technology to investigate the existence of global gene expression patterns in individuals with BD	-Differently expressed genes were observed during iPSC to NPC differentiation in BD patients -Enrichment of genes correlated with cell cycle regulation and homeostasis	Kim et al. (2015)
Exploratory	N/a	iPSCs Neurons NPCs	A family-based paradigm model was used to examine iPSCs lines from brothers with DB compared to non-BD parents	-NPCs and neuronal cells derived from BD patients showed phenotypical differences in channeled subunits and Wnt pathway components related to neuroplasticity	Madison et al. (2015)
Exploratory and Treatment	Lithium	iPSC-derived neurons	Study of phenotypical differences in neurons derived from iPSCs generated from lithium responder and non-responder patients and controls	- Discovered that BD derived neurons were hyperexcitable compared to control neurons -Lithium reversed hyperexcitability only on neurons derived from lithium responders	Mertens et al. (2015, 2016)
Treatment	Lithium	iPSC-derived neurons	Study of neurons derived from BD lithium responder patients to gain insight to BD molecular mechanisms and lithium targets	-Lithium acts on CRMP2 which alters neuronal cytoskeletal dynamics and spine formation in BD iPSC derived neurons	Tobe et al. (2017)
Exploratory	N/a	iPSCs NPCs	Examination of transcriptomes during the transition from iPSCs to NPCs in BD patients and controls using RNA sequencing	-Increased expression of genes related to inflammation in BD-derived NPCs -The most significantly differentially expressed genes were NLRP2 and genes associated with GABA and dopamine receptor canonical pathways	Vizlin-Hodzic et al. (2017)
Exploratory	N/a	iPSC-derived neurons	Modeled and compared activity of neurons derived iPSCs from lithium responders and non-responders BD patients	-Created a model to predict if a patient will respond to lithium or not with a 92% accuracy rate	Stern et al. (2018)
Exploratory	N/a	iPSC-derived neurons	Study of the behavior of neurons of hippocampus CA3 and dentate gyrus and motor neurons from lithium responders and non-responders BD patients	- Hyperexcitability is dependent on the type of neurons and unstable in neurons derived from lithium non-responders	Stern et al. (2020a,b)
Exploratory	N/a	Organoids	Exploration of functional mechanisms in BD by comparing BD-derived and control organoids	-BD derived patient neurons expressed upregulation of genes involved in neurogenesis, cell adhesion, synaptic morphology, and function and a downregulation of genes involved in immune signaling along with a reduction in mitochondria-associated endoplasmic reticulum membranes (MAMs) -BD derived organoids responded differently to electrical stimuli but similarly to non-BD derived models in the absence of electrical stimuli	Kathuria et al. (2020)
Treatment	Lithium	iPSC-derived NPCs and lymphoblastoid cell lines	Examination of cellular phenotypes of lithium responder and non-responder BD patients compared to healthy controls	-In vitro lithium treatment of BD derived iPSC lines encouraged cell proliferation, enhanced the expression of BCL2 and GSK3B and lowered mitochondrial membrane potential (MMP) - Low MMP was reversed using in vitro lithium exposure but remained unaffected in clinical lithium therapy treatment	Paul et al. (2020)
Treatment	Valproate	iPSCs	Examination of how valproate interacts with iPSCs from lithium responders and non-responders	-Valproate rescued dysregulation of cell proliferation and cell death in lithium responders and non-responders	Paul et al. (2020)
Exploratory	N/a	iPSC-derived astrocytes	Generated astrocyte models derived from iPSC from BD patients and controls to examine inflammation related phenotypes	-IL-6, a proinflammatory cytokine, was upregulated in iPSC-derived astrocytes from BD patients compared to controls -When BD iPSC derived astrocytes were co-cultured with neurons, there was a reduction in neuronal activity	Vadodaria et al. (2021)
Exploratory and Treatment	Lithiumand Valproic Acid	iPSC-derived neurons	Examined lithium resistance in iPSC-derived neurons derived from lithium non-responder patients with B	-NR neurons displayed less activity of the Wnt/β-catenin signaling pathway and a significant decrease in LEF1 expression	Santos et al. (2021)
Exploratory	N/a	Stem Cells Neurons NPCs	Mapped the presence and arrangement of lithium ions in cells	-Unveiled the mechanisms of lithium ion distribution in multiple cell types including patient derived cells from individuals with BD to further explore and understand lithium- response pathways	McGhee et al. (2021)
Exploratory	N/a	iPSC-derived neurons	Examined the circadian-rhythm regulation in neurons derived from iPSCs of BD lithium responder and non-responder patients	- BD neurons showed differences in circadian-rhythm regulation with the most significant difference seen in lithium non-responders	Mishra et al. (2021)
Treatment	Trimetazidine	iPSC-derived neurons and astrocytes	Study aimed to repurpose trimetazidine to treat bipolar depression	- Trimetazidine increased the production of ATP, which can be deficient in BD patients, by altering cellular metabolic processes	Bortolasci et al. (2023)
Treatment	Lithium	iPSC-derived cortical spheroids	Examination of phenotypic effects of prolonged lithium exposure on corticoid spheroids derived from BD patients	-BD patient-derived corticoid spheroids exhibited transcriptional profiles with enriched differentially expressed genes associated with kidney function and ion homeostasis when exposed to chronic lithium treatment	Osete et al. (2023)
Treatment	Apigenin	iPSC-derived astrocytes	Tested the effects of apigenin, a neuroprotective and anti-inflammatory compound, on stimulated iPSC-derived astrocytes from BD and control individuals to examine alternative treatment methods for bipolar disorder	-BD and control astrocytes experienced an increase in the percentage of IL-6 expressing astrocytes irrespective of the proinflammatory stimuli used (IL-1b or TNF-a) - Both BD and control astrocytes showed higher sensitivity to proinflammatory cytokine IL-6 compared to TNF-a -Apigenin treatment successfully counteracted the inflammatory response in BD and control astrocytes	This study

Study Type	Treatment	Cell/ Tissue Type(s)	Involvement of iPSCs in Study	Findings	Reference
Exploratory and Treatment	Sertraline	HPCs	Exploratory: To examine the effects of chronic stress, a known cofounder of depression, researchers exposed HPCs from healthy individuals to cortisol and dexamethasone (or dex, synthetic cortisol) Treatment: The HPCs were then treated with sertraline, the most prescribed SSRI for MDD	Exploratory: -Healthy HPCs exposed to dex and cortisol exhibited a decrease in cell proliferation and neuronal differentiation -Increased expression of genes involved in cell cycle inhibition Treatment: -Increase in glucocorticoid receptor transcription rate -Alterations in neurogenesis associated gene expression profiles	Anacker et al. (2011)
Exploratory	N/a	HPCs	This study explored the effects of proinflammatory cytokine interleukin-1b (IL-1b) on neuronal generation	-Increase in transcripts for kynurenine 3-monooxygenase (KMO) enzyme during cell differentiation -Blocking KMO resulted in the restoration reduced neurogenesis caused by IL-1b exposure	Zunszain et al. (2012)
Exploratory	N/a	HPCs	Examined the effects of cortisol exposure in MDD vs non-MDD hippocampal progenitor cells	-Increase in cell proliferation -Decrease in neurogenesis	Anacker et al. (2013a)
Treatment	Glucocorticoid regulated kinase 1 (SKG1)	HPCs	Explored the effects of glucocorticoid regulated kinase 1 (SKG1) on HPCs following cortisol exposure	-The effects of cortisol exposure in HPCs can be mitigated by SKG1	Anacker et al. (2013b)
Treatment	Sertraline	hAD-SCs	Examined the changes in human adipose-derived stem cells after sertraline treatment	-Increase in proliferation	Razavi et al. (2014)
Treatment	Venlafaxine (SNRI) Eicosapentaenoic acid (EPA) Sertraline (SSRI) docosahexaenoic acid (DHA)	HPCs	HPCs were treated with IL-1b to induce inflammation followed by exposure of various forms of treatment for MDD to examine the difference in molecular mechanisms by treatment	-Venlafaxine and eicosapentaenoic acid had anti-inflammatory effects HPCs exposed to IL-1b -Sertraline and docosahexaenoic acid had pro-inflammatory effects on IL-1b treated HPCs -All treatments were associated with a decrease in NF-kB pathway activity	Horowitz et al. (2015)
Treatment	Paroxetine	Neurons	Explored the effects of paroxetine neuronal differentiation and cell proliferation on adipose-derived stem cells	-Paroxetine enhanced neurogenic differentiation and proliferation rate	Jahromi et al. (2016)
Treatment	IL-1b	HPCs	Treated HPCs with IL-1b to examine effects on the kynurenic pathway	-IL-1b exposure lead to a restoration of neurogenesis	Borsini et al. (2017)
Exploratory	N/a	HPCs	The effects of interferon-a to investigate the mechanisms of inflammation-induced depression	-Reduction in neurogenesis -Increased cell death -Increase in expression of ubiquitin-specific peptidase 18 (USP18) and interferon-stimulated gene 15 (ISG15) was responsible for the reduction of neurogenesis -Increased expression in interleukin-6 (IL-6) is responsible for the increase in cell death	Borsini et al. (2018)
Treatment	Ketamine	Neurons	Examined the effects of ketamine on prefrontal hippocampal neurons derived from MDD patients	-Ketamine increased structural plasticity in MDD derived dopaminergic neurons	Cavalleri et al. (2018)
Exploratory	Ketamine	Neurons	Examined current literature on ketamine exposure on iPSCs to treat MDD	-Glutamate burst induced structural plasticity	Collo and Merlo Pitch (2018)
Exploratory	N/a	HPCs	Exposure of serum derived from patients with MDD on HPCs from non-MDD study participants	-Treated HPCs showed an increase in cell apoptosis and a decrease in neurogenesis matching that of MDD patient derived HPCs -Increase in apoptosis and decrease in neurogenesis exacerbated following interferon-a treatment	Borsini et al. (2019)
Treatment	Ropinirole and Pramipexole	Neurons	Examined the effect of ropinirole and pramipexole on neurons derived from patients with treatment resistant depression	-Ropinirole and pramipexole regulated structural plasticity in neurons derived from treatment resistant depression patients	Collo et al. (2018b)
Treatment	Serotonin and Latuda	Neurons	Treatment (Serotonin): Exposed neurons derived from non-SSRI responders to Serotonin to examine changes in activity Treatment (Latuda): Exposed SSRI non-responders to Latuda, an antipsychotic drug) to examine changes in activity	-Serotonin: Increase in activity related to the upregulation of excitatory serotonergic receptors (5-HT7 and 5-HT2A), -Latuda rescued hyperactivity in SSRI non responders	Vadodaria et al. (2019a)
Exploratory and Treatment		iPSC-derived serotonergic neurons	Exploratory: iPSCs from SSRI responders, non-responders and individuals with no history of MDD were juxtaposed to study differences in morphology, action and behavior Treatment: Treated cells with SSRIs to study the differences in the expression of serotonergic genes between the controlled and patient groups	-Exploratory: No difference found in the expression of serotonergic genes -Treatment: In SSRI non-responders, the genes protocadherin alpha 6 and alpha 8 exhibited were differentially expressed compared to SSRI responders	Vadodaria et al. (2019b)
Exploratory	N/a	HPCs	Examined the effects of IL-6 on HPCs from MDD and non-MDD derived HPCs	-Pro and anti-inflammatory effects on HPCs based on levels and presence of other cytokines	Borsini et al. (2020)
Treatment	Paroxetine	Oligodendrocytes	Examined the risk of neurotoxicity caused by SSRI medication; iPSC derived oligodendrocytes were exposed to MDD treatment paroxetine	-Reduction in neurite outgrowth -Cell population decrease	Zhong et al. (2020)
Treatment	Bupropion	Cortical neurons	Examined the effects of bupropion on cortical neurons derived from MDD bupropion responders compared to a controlled group	-Cortical neurons derived from MDD patients showed a difference in synaptic connection, gene expression and morphology	Avior et al. (2021)
Exploratory	N/a	Glial cells	Explored the effects of chronic cortisol exposure on glial cells from MDD patients	-Differentially expressed genes related to ion homeostasis, G-protein-coupled receptors (GPCR), and synaptic signaling	Heard et al. (2021)
Exploratory	N/a	Neurons	Examined neurons derived from MDD patients and compared them to a control	-MDD patient derived neurons demonstrated functional and bioenergetic differences -Increased electrical activity -Change in sodium ion channels	Triebelhorn et al. (2022)
Exploratory	N/a	Forebrain organoids	Examined the difference in morphology, behavior, and function of corticoid organoids derived from MDD patients with a history of attempted suicide	-GABAergic interneurons and ventral forebrain organoids form MDD patients exhibited an increase in neuronal firing -Difference in neuronal morphology -Decrease in calcium signals -The dysregulation in neuronal morphology and behavior may be due to the decreased expression of 5-HT2C, a serotonergic receptor	Lu et al. (2023)

[TFN1] Abbreviations: 3D: three-dimensional, ATP: adenosine triphosphate, BCL2: B-cell lymphoma 2, BD: bipolar disorder, CRMP2: collapsin response mediator protein-2, DEGs: differentially expressed genes, GABA: gamma-aminobutyric acid, GSK3: glycogen synthase kinase-3, hsa-miR: human microRNAs, IL-1b: interleukin-1-beta, IL-6: interleukin-6, iPSCs: induced pluripotent stem cells, LEF1: lymphoid enhancer-binding factor 1, Li: lithium, MAMs: mitochondria-associated endoplasmic reticulum membranes, MMP: mitochondrial membrane potential, NLRP2: NLR family pyrin domain containing 2, NPCs: neuronal progenitor cells, NR: non-responder, RNA: ribonucleic acid, TNF-a: tumor necrosis factor alpha, Wnt: wingless-related integration site