Single-Cell RNA Sequencing of Human Pluripotent Stem Cell-Derived Macrophages for Quality Control of The Cell Therapy Product

Overview

Journal Front Genet

Date 2022 Feb 17

PMID 35173760

Authors

Hye-Yeong Jo

Hyang-Hee Seo

Dayeon Gil

Youngchan Park

Hyeong-Jun Han

Hyo-Won Han

Rajesh K Thimmulappa

Sang Cheol Kim

Jung-Hyun Kim

Affiliations

Soon will be listed here.

Abstract

Macrophages exhibit high plasticity to achieve their roles in maintaining tissue homeostasis, innate immunity, tissue repair and regeneration. Therefore, macrophages are being evaluated for cell-based therapeutics against inflammatory disorders and cancer. To overcome the limitation related to expansion of primary macrophages and cell numbers, human pluripotent stem cell (hPSC)-derived macrophages are considered as an alternative source of primary macrophages for clinical application. However, the quality of hPSC-derived macrophages with respect to the biological homogeneity remains still unclear. We previously reported a technique to produce hPSC-derived macrophages referred to as iMACs, which is amenable for scale-up. In this study, we have evaluated the biological homogeneity of the iMACs using a transcriptome dataset of 6,230 iMACs obtained by single-cell RNA sequencing. The dataset provides a valuable genomic profile for understanding the molecular characteristics of hPSC-derived macrophage cells and provide a measurement of transcriptomic homogeneity. Our study highlights the usefulness of single cell RNA-seq data in quality control of the cell-based therapy products.

Citing Articles

Deep skin fibroblast-mediated macrophage recruitment supports acute wound healing.

Amuso V, Haas M, Cooper P, Chatterjee R, Hafiz S, Salameh S bioRxiv. 2024; .

PMID: 39149286 PMC: 11326280. DOI: 10.1101/2024.08.09.607357.

Generation of chimeric antigen receptor macrophages from human pluripotent stem cells to target glioblastoma.

Jin G, Chang Y, Bao X Immunooncol Technol. 2024; 20:100409.

PMID: 38192614 PMC: 10772262. DOI: 10.1016/j.iotech.2023.100409.

Recent Approaches to Design and Analysis of Electrical Impedance Systems for Single Cells Using Machine Learning.

Ferguson C, Zhang Y, Palego C, Cheng X Sensors (Basel). 2023; 23(13).

PMID: 37447838 PMC: 10346645. DOI: 10.3390/s23135990.

Single-Cell Transcriptome Analysis Identifies Subclusters with Inflammatory Fibroblast Responses in Localized Scleroderma.

Werner G, Sanyal A, Mirizio E, Hutchins T, Tabib T, Lafyatis R Int J Mol Sci. 2023; 24(12).

PMID: 37372943 PMC: 10298454. DOI: 10.3390/ijms24129796.

iPSC-Derived Macrophages: The Differentiation Protocol Affects Cell Immune Characteristics and Differentiation Trajectories.

Klepikova A, Nenasheva T, Sheveleva O, Protasova E, Antonov D, Gainullina A Int J Mol Sci. 2022; 23(24).

PMID: 36555728 PMC: 9781144. DOI: 10.3390/ijms232416087.

References

Das A, Sinha M, Datta S, Abas M, Chaffee S, Sen C . Monocyte and macrophage plasticity in tissue repair and regeneration. Am J Pathol. 2015; 185(10):2596-606. PMC: 4607753. DOI: 10.1016/j.ajpath.2015.06.001. View

Macosko E, Basu A, Satija R, Nemesh J, Shekhar K, Goldman M . Highly Parallel Genome-wide Expression Profiling of Individual Cells Using Nanoliter Droplets. Cell. 2015; 161(5):1202-1214. PMC: 4481139. DOI: 10.1016/j.cell.2015.05.002. View

Kim J, Jo H, Ha H, Kim Y . Korea National Stem Cell Bank. Stem Cell Res. 2021; 53:102270. DOI: 10.1016/j.scr.2021.102270. View

Zhang L, Tian L, Dai X, Yu H, Wang J, Lei A . Pluripotent stem cell-derived CAR-macrophage cells with antigen-dependent anti-cancer cell functions. J Hematol Oncol. 2020; 13(1):153. PMC: 7656711. DOI: 10.1186/s13045-020-00983-2. View

Fidler I . Inhibition of pulmonary metastasis by intravenous injection of specifically activated macrophages. Cancer Res. 1974; 34(5):1074-8. View

Wynn T, Chawla A, Pollard J . Macrophage biology in development, homeostasis and disease. Nature. 2013; 496(7446):445-55. PMC: 3725458. DOI: 10.1038/nature12034. View

Biswas S, Mantovani A . Macrophage plasticity and interaction with lymphocyte subsets: cancer as a paradigm. Nat Immunol. 2010; 11(10):889-96. DOI: 10.1038/ni.1937. View

Powell R, Marston W, Berceli S, Guzman R, Henry T, Longcore A . Cellular therapy with Ixmyelocel-T to treat critical limb ischemia: the randomized, double-blind, placebo-controlled RESTORE-CLI trial. Mol Ther. 2012; 20(6):1280-6. PMC: 3369291. DOI: 10.1038/mt.2012.52. View

Alvey C, Discher D . Engineering macrophages to eat cancer: from "marker of self" CD47 and phagocytosis to differentiation. J Leukoc Biol. 2017; 102(1):31-40. PMC: 6608056. DOI: 10.1189/jlb.4RI1216-516R. View

10.

Smolander J, Junttila S, Venalainen M, Elo L . ILoReg: a tool for high-resolution cell population identification from single-cell RNA-seq data. Bioinformatics. 2020; 37(8):1107-1114. PMC: 8150131. DOI: 10.1093/bioinformatics/btaa919. View

11.

Andreesen R, Hennemann B . Adoptive immunotherapy with autologous macrophages: current status and future perspectives. Pathobiology. 1991; 59(4):259-63. DOI: 10.1159/000163658. View

12.

Lee S, Kivimae S, Dolor A, Szoka F . Macrophage-based cell therapies: The long and winding road. J Control Release. 2016; 240:527-540. PMC: 5064880. DOI: 10.1016/j.jconrel.2016.07.018. View

13.

Kunisch E, Fuhrmann R, Roth A, Winter R, Lungershausen W, Kinne R . Macrophage specificity of three anti-CD68 monoclonal antibodies (KP1, EBM11, and PGM1) widely used for immunohistochemistry and flow cytometry. Ann Rheum Dis. 2004; 63(7):774-84. PMC: 1755048. DOI: 10.1136/ard.2003.013029. View

14.

Thomson J, Itskovitz-Eldor J, Shapiro S, Waknitz M, Swiergiel J, Marshall V . Embryonic stem cell lines derived from human blastocysts. Science. 1998; 282(5391):1145-7. DOI: 10.1126/science.282.5391.1145. View

15.

Starkey Lewis P, Moroni F, Forbes S . Macrophages as a Cell-Based Therapy for Liver Disease. Semin Liver Dis. 2019; 39(4):442-451. DOI: 10.1055/s-0039-1688502. View

16.

Alvey C, Spinler K, Irianto J, Pfeifer C, Hayes B, Xia Y . SIRPA-Inhibited, Marrow-Derived Macrophages Engorge, Accumulate, and Differentiate in Antibody-Targeted Regression of Solid Tumors. Curr Biol. 2017; 27(14):2065-2077.e6. PMC: 5846676. DOI: 10.1016/j.cub.2017.06.005. View

17.

Hu M, Walmsley G, Barnes L, Weiskopf K, Rennert R, Duscher D . Delivery of monocyte lineage cells in a biomimetic scaffold enhances tissue repair. JCI Insight. 2017; 2(19). PMC: 5841872. DOI: 10.1172/jci.insight.96260. View

18.

Sharkey J, Ressel L, Brillant N, Scarfe L, Wilm B, Park B . A Noninvasive Imaging Toolbox Indicates Limited Therapeutic Potential of Conditionally Activated Macrophages in a Mouse Model of Multiple Organ Dysfunction. Stem Cells Int. 2019; 2019:7386954. PMC: 6466849. DOI: 10.1155/2019/7386954. View

19.

Han H, Seo H, Jo H, Han H, Falcao V, Delorme V . Drug Discovery Platform Targeting M. tuberculosis with Human Embryonic Stem Cell-Derived Macrophages. Stem Cell Reports. 2019; 13(6):980-991. PMC: 6915848. DOI: 10.1016/j.stemcr.2019.10.002. View

20.

Mosser D, Edwards J . Exploring the full spectrum of macrophage activation. Nat Rev Immunol. 2008; 8(12):958-69. PMC: 2724991. DOI: 10.1038/nri2448. View