Modeling HIV-1 neuropathogenesis using three-dimensional human brain organoids (hBORGs) with HIV-1 infected microglia

Reis, Roberta Sena; Sant, Shilpa; Keeney, Hannah; Wagner, M.; Ayyavoo, Velpandi

doi:10.1038/s41598-020-72214-0

Cited by 73 publications

(63 citation statements)

References 86 publications

(141 reference statements)

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“…Microtumors of 150 µm (referred to as ‘small’ microtumors) and 600 µm diameters (referred to as ‘large’ microtumors) were obtained by seeding T47D cells in non-adhesive polyethylene glycol (PEG) hydrogel microwell devices, as previously described [ 16 , 17 , 29 , 33 ]. Briefly, 1 × 1 cm 2 microwell devices were fabricated using polydimethyl siloxane (PDMS) stamps containing posts of either 150 µm or 600 µm in diameter with equal height.…”

Section: Methodsmentioning

confidence: 99%

Identifying Molecular Signatures of Distinct Modes of Collective Migration in Response to the Microenvironment Using Three-Dimensional Breast Cancer Models

Ardila

Aggarwal

Singh

et al. 2021

Cancers

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Collective cell migration is a key feature of transition of ductal carcinoma in situ (DCIS) to invasive ductal carcinoma (IDC) among many other cancers, yet the microenvironmental factors and underlying mechanisms that trigger collective migration remain poorly understood. Here, we investigated two microenvironmental factors, tumor-intrinsic hypoxia and tumor-secreted factors (secretome), as triggers of collective migration using three-dimensional (3D) discrete-sized microtumor models that recapitulate hallmarks of DCIS-IDC transition. Interestingly, the two factors induced two distinct modes of collective migration: directional and radial migration in the 3D microtumors generated from the same breast cancer cell line model, T47D. Without external stimulus, large (600 µm) T47D microtumors exhibited tumor-intrinsic hypoxia and directional migration, while small (150 µm), non-hypoxic microtumors exhibited radial migration only when exposed to the secretome of large microtumors. To investigate the mechanisms underlying hypoxia- and secretome-induced directional vs. radial migration modes, we performed differential gene expression analysis of hypoxia- and secretome-induced migratory microtumors compared with non-hypoxic, non-migratory small microtumors as controls. We propose unique gene signature sets related to tumor-intrinsic hypoxia, hypoxia-induced epithelial-mesenchymal transition (EMT), as well as hypoxia-induced directional migration and secretome-induced radial migration. Gene Set Enrichment Analysis (GSEA) and protein-protein interaction (PPI) network analysis revealed enrichment and potential interaction between hypoxia, EMT, and migration gene signatures for the hypoxia-induced directional migration. In contrast, hypoxia and EMT were not enriched in the secretome-induced radial migration, suggesting that complete EMT may not be required for radial migration. Survival analysis identified unique genes associated with low survival rate and poor prognosis in TCGA-breast invasive carcinoma dataset from our tumor-intrinsic hypoxia gene signature (CXCR4, FOXO3, LDH, NDRG1), hypoxia-induced EMT gene signature (EFEMP2, MGP), and directional migration gene signature (MAP3K3, PI3K3R3). NOS3 was common between hypoxia and migration gene signature. Survival analysis from secretome-induced radial migration identified ATM, KCNMA1 (hypoxia gene signature), and KLF4, IFITM1, EFNA1, TGFBR1 (migration gene signature) to be associated with poor survival rate. In conclusion, our unique 3D cultures with controlled microenvironments respond to different microenvironmental factors, tumor-intrinsic hypoxia, and secretome by adopting distinct collective migration modes and their gene expression analysis highlights the phenotypic heterogeneity and plasticity of epithelial cancer cells.

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Section: Methodsmentioning

confidence: 99%

Identifying Molecular Signatures of Distinct Modes of Collective Migration in Response to the Microenvironment Using Three-Dimensional Breast Cancer Models

Ardila

Aggarwal

Singh

et al. 2021

Cancers

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“…However, microglial cells and endothelial cells are largely absent in original CO systems. Several protocols have been developed to co-culture iPSC-derived microglia with CO or iPSC-derived neural cells (Dos Reis et al, 2020 ; Fritsche et al, 2020 ; Hasselmann and Blurton-Jones, 2020 ; Tanaka and Park, 2021 ). While these methods are effective at generating MCOs, they require separate culturing systems with multiple additional steps as well as optimization of cell number ratio.…”

Section: Discussionmentioning

confidence: 99%

“…However, some of these mechanisms have not fully been explored during early neurodevelopment, but MCOs may present a unique model to study these critical interactions, particularly in the context of elucidating mechanisms underlying neurodevelopmental disorders and neurodegenerative diseases. Furthermore, as microglia are known to play critical roles in inflammation and infection, MCOs provide an excellent model for neurotropic viral infections such as HIV and ZIKV (Muffat et al, 2018 ; Dos Reis et al, 2020 ; Qiao et al, 2020 ; Bodnar et al, 2021 ). While organoids have been used to study viral infections before (Cugola et al, 2016 ; Dang et al, 2016 ; Qian et al, 2016 ; Gabriel et al, 2017 ; Li et al, 2017b ; Watanabe et al, 2017 ), these models did not include microglia, which are critical to the immune response.…”

Section: Discussionmentioning

confidence: 99%

“…Optimization of the heparin concentration or other regulators may improve the generation of microglia in MCO. Another strategy might be the co-culture of iPSC-derived microglia with the CO (Dos Reis et al, 2020 ; Rai et al, 2020 ), which may increase the number of microglia cells in CO, although the process does not mimic normal brain development. Another limit of our MCO model is that there are still several cell types missing, including endothelial cells/vasculature.…”

Section: Discussionmentioning

confidence: 99%

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Novel Scalable and Simplified System to Generate Microglia-Containing Cerebral Organoids From Human Induced Pluripotent Stem Cells

Bodnar

Zhang

Liu

et al. 2021

Front. Cell. Neurosci.

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Human cerebral organoid (CO) is a three-dimensional (3D) cell culture system that recapitulates the developing human brain. While CO has proved an invaluable tool for studying neurological disorders in a more clinically relevant matter, there have still been several shortcomings including CO variability and reproducibility as well as lack of or underrepresentation of certain cell types typically found in the brain. As the technology to generate COs has continued to improve, more efficient and streamlined protocols have addressed some of these issues. Here we present a novel scalable and simplified system to generate microglia-containing CO (MCO). We characterize the cell types and dynamic development of MCOs and validate that these MCOs harbor microglia, astrocytes, neurons, and neural stem/progenitor cells, maturing in a manner that reflects human brain development. We introduce a novel technique for the generation of embryoid bodies (EBs) directly from induced pluripotent stem cells (iPSCs) that involves simplified steps of transitioning directly from 3D cultures as well as orbital shaking culture in a standard 6-well culture plate. This allows for the generation of MCOs with an easy-to-use system that is affordable and accessible by any general lab.

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“… 1 References: Immune deficiency virus (HIV): [ 112 , 117 , 124 , 197 , 198 , 199 , 200 , 201 ]. Measles virus: [ 135 , 202 , 203 ].…”

Section: Tablementioning

confidence: 99%

The Influence of Virus Infection on Microglia and Accelerated Brain Aging

Filgueira

Larionov

Lannes

2021

Cells

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Microglia are the resident immune cells of the central nervous system contributing substantially to health and disease. There is increasing evidence that inflammatory microglia may induce or accelerate brain aging, by interfering with physiological repair and remodeling processes. Many viral infections affect the brain and interfere with microglia functions, including human immune deficiency virus, flaviviruses, SARS-CoV-2, influenza, and human herpes viruses. Especially chronic viral infections causing low-grade neuroinflammation may contribute to brain aging. This review elucidates the potential role of various neurotropic viruses in microglia-driven neurocognitive deficiencies and possibly accelerated brain aging.

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Modeling HIV-1 neuropathogenesis using three-dimensional human brain organoids (hBORGs) with HIV-1 infected microglia

Cited by 73 publications

References 86 publications

Identifying Molecular Signatures of Distinct Modes of Collective Migration in Response to the Microenvironment Using Three-Dimensional Breast Cancer Models

Identifying Molecular Signatures of Distinct Modes of Collective Migration in Response to the Microenvironment Using Three-Dimensional Breast Cancer Models

Novel Scalable and Simplified System to Generate Microglia-Containing Cerebral Organoids From Human Induced Pluripotent Stem Cells

The Influence of Virus Infection on Microglia and Accelerated Brain Aging

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