Improving hematopoietic differentiation from human induced pluripotent stem cells by the modulation of Hippo signaling with a diarylheptanoid derivative.

Diarylheptanoid Hippo signaling pathway Human induced pluripotent stem cells Primitive hematopoietic stem and progenitor cells

Journal

Stem cell research & therapy
ISSN: 1757-6512
Titre abrégé: Stem Cell Res Ther
Pays: England
ID NLM: 101527581

Informations de publication

Date de publication:
03 Mar 2024
Historique:
received: 04 07 2023
accepted: 27 02 2024
medline: 4 3 2024
pubmed: 4 3 2024
entrez: 3 3 2024
Statut: epublish

Résumé

The diarylheptanoid ASPP 049 has improved the quality of adult hematopoietic stem cell (HSC) expansion ex vivo through long-term reconstitution in animal models. However, its effect on hematopoietic regeneration from human induced pluripotent stem cells (hiPSCs) is unknown. We utilized a defined cocktail of cytokines without serum or feeder followed by the supplementation of ASPP 049 to produce hematopoietic stem/progenitor cells (HSPCs). Flow cytometry and trypan blue exclusion analysis were used to identify nonadherent and adherent cells. Nonadherent cells were harvested to investigate the effect of ASPP 049 on multipotency using LTC-IC and CFU assays. Subsequently, the mechanism of action was explored through transcriptomic profiles, which were validated by qRT-PCR, immunoblotting, and immunofluorescence analysis. The supplementation of ASPP 049 increased the number of phenotypically defined primitive HSPCs (CD34 These findings suggest that ASPP 049 can improve HSC-generating protocols with proliferative potentials, self-renewal ability, unbiased differentiation, and a definable mechanism of action for the clinical perspective of hematopoietic regenerative medicine.

Sections du résumé

BACKGROUND BACKGROUND
The diarylheptanoid ASPP 049 has improved the quality of adult hematopoietic stem cell (HSC) expansion ex vivo through long-term reconstitution in animal models. However, its effect on hematopoietic regeneration from human induced pluripotent stem cells (hiPSCs) is unknown.
METHOD METHODS
We utilized a defined cocktail of cytokines without serum or feeder followed by the supplementation of ASPP 049 to produce hematopoietic stem/progenitor cells (HSPCs). Flow cytometry and trypan blue exclusion analysis were used to identify nonadherent and adherent cells. Nonadherent cells were harvested to investigate the effect of ASPP 049 on multipotency using LTC-IC and CFU assays. Subsequently, the mechanism of action was explored through transcriptomic profiles, which were validated by qRT-PCR, immunoblotting, and immunofluorescence analysis.
RESULT RESULTS
The supplementation of ASPP 049 increased the number of phenotypically defined primitive HSPCs (CD34
CONCLUSION CONCLUSIONS
These findings suggest that ASPP 049 can improve HSC-generating protocols with proliferative potentials, self-renewal ability, unbiased differentiation, and a definable mechanism of action for the clinical perspective of hematopoietic regenerative medicine.

Identifiants

pubmed: 38433217
doi: 10.1186/s13287-024-03686-4
pii: 10.1186/s13287-024-03686-4
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

60

Subventions

Organisme : Mahidol University
ID : MU's Strategic Research Fund: fiscal year 2023
Organisme : National Research Council of Thailand
ID : N41A660154
Organisme : Ministry of Science and Technology of Thailand
ID : the Science Achievement Scholarship of Thailand
Organisme : Faculty of Science, Mahidol University
ID : the Central Instrument Facility (CIF)

Informations de copyright

© 2024. The Author(s).

Références

Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, Tomoda K, et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell. 2007;131(5):861–72.
pubmed: 18035408 doi: 10.1016/j.cell.2007.11.019
Rowe RG, Daley GQ. Induced pluripotent stem cells in disease modelling and drug discovery. Nat Rev Genet. 2019;20(7):377–88.
pubmed: 30737492 pmcid: 6584039 doi: 10.1038/s41576-019-0100-z
Hansen M, von Lindern M, van den Akker E, Varga E. Human-induced pluripotent stem cell-derived blood products: state of the art and future directions. FEBS Lett. 2019;593(23):3288–303.
pubmed: 31520530 doi: 10.1002/1873-3468.13599
Vo LT, Daley GQ. De novo generation of HSCs from somatic and pluripotent stem cell sources. Blood. 2015;125(17):2641–8.
pubmed: 25762177 pmcid: 4408289 doi: 10.1182/blood-2014-10-570234
Sugimura R, Jha DK, Han A, Soria-Valles C, da Rocha EL, Lu YF, et al. Haematopoietic stem and progenitor cells from human pluripotent stem cells. Nature. 2017;545(7655):432–8.
pubmed: 28514439 pmcid: 5872146 doi: 10.1038/nature22370
Doulatov S, Vo LT, Chou SS, Kim PG, Arora N, Li H, et al. Induction of multipotential hematopoietic progenitors from human pluripotent stem cells via respecification of lineage-restricted precursors. Cell Stem Cell. 2013;13(4):459–70.
pubmed: 24094326 doi: 10.1016/j.stem.2013.09.002
Gori JL, Butler JM, Chan YY, Chandrasekaran D, Poulos MG, Ginsberg M, et al. Vascular niche promotes hematopoietic multipotent progenitor formation from pluripotent stem cells. J Clin Invest. 2015;125(3):1243–54.
pubmed: 25664855 pmcid: 4362238 doi: 10.1172/JCI79328
Kennedy M, Awong G, Sturgeon CM, Ditadi A, LaMotte-Mohs R, Zuniga-Pflucker JC, et al. T lymphocyte potential marks the emergence of definitive hematopoietic progenitors in human pluripotent stem cell differentiation cultures. Cell Rep. 2012;2(6):1722–35.
pubmed: 23219550 doi: 10.1016/j.celrep.2012.11.003
Sturgeon CM, Ditadi A, Awong G, Kennedy M, Keller G. Wnt signaling controls the specification of definitive and primitive hematopoiesis from human pluripotent stem cells. Nat Biotechnol. 2014;32(6):554–61.
pubmed: 24837661 pmcid: 4152856 doi: 10.1038/nbt.2915
Choi KD, Vodyanik MA, Togarrati PP, Suknuntha K, Kumar A, Samarjeet F, et al. Identification of the hemogenic endothelial progenitor and its direct precursor in human pluripotent stem cell differentiation cultures. Cell Rep. 2012;2(3):553–67.
pubmed: 22981233 pmcid: 3462245 doi: 10.1016/j.celrep.2012.08.002
Lacaud G, Kouskoff V. Hemangioblast, hemogenic endothelium, and primitive versus definitive hematopoiesis. Exp Hematol. 2017;49:19–24.
pubmed: 28043822 doi: 10.1016/j.exphem.2016.12.009
Slukvin I. Hematopoietic specification from human pluripotent stem cells current advances and challenges toward de novo generation of hematopoietic stem cells. Blood. 2013;122:25.
doi: 10.1182/blood-2013-07-474825
Canu G, Ruhrberg C. First blood: the endothelial origins of hematopoietic progenitors. Angiogenesis. 2021;24(2):199–211.
pubmed: 33783643 pmcid: 8205888 doi: 10.1007/s10456-021-09783-9
Lange L, Morgan M, Schambach A. The hemogenic endothelium: a critical source for the generation of PSC-derived hematopoietic stem and progenitor cells. Cell Mol Life Sci. 2021;78(9):4143–60.
pubmed: 33559689 pmcid: 8164610 doi: 10.1007/s00018-021-03777-y
Demirci S, Leonard A, Tisdale JF. Hematopoietic stem cells from pluripotent stem cells: clinical potential, challenges, and future perspectives. Stem Cells Transl Med. 2020;9(12):1549–57.
pubmed: 32725882 pmcid: 7695636 doi: 10.1002/sctm.20-0247
Shen J, Xu Y, Zhang S, Lyu S, Huo Y, Zhu Y, et al. Single-cell transcriptome of early hematopoiesis guides arterial endothelial-enhanced functional T cell generation from human PSCs. Sci Adv. 2021;7(36):eabi9787.
pubmed: 34516916 pmcid: 8442917 doi: 10.1126/sciadv.abi9787
Grigoriadis AE, Kennedy M, Bozec A, Brunton F, Stenbeck G, Park IH, et al. Directed differentiation of hematopoietic precursors and functional osteoclasts from human ES and iPS cells. Blood. 2010;115(14):2769–76.
pubmed: 20065292 pmcid: 2854424 doi: 10.1182/blood-2009-07-234690
Galat Y, Dambaeva S, Elcheva I, Khanolkar A, Beaman K, Iannaccone PM, et al. Cytokine-free directed differentiation of human pluripotent stem cells efficiently produces hemogenic endothelium with lymphoid potential. Stem Cell Res Ther. 2017;8(1):67.
pubmed: 28302184 pmcid: 5356295 doi: 10.1186/s13287-017-0519-0
Tan YT, Ye L, Xie F, Beyer AI, Muench MO, Wang J, et al. Respecifying human iPSC-derived blood cells into highly engraftable hematopoietic stem and progenitor cells with a single factor. Proc Natl Acad Sci USA. 2018;115(9):2180–5.
pubmed: 29386396 pmcid: 5834708 doi: 10.1073/pnas.1718446115
Li X, Xia C, Wang T, Liu L, Zhao Q, Yang D, et al. Pyrimidoindole derivative UM171 enhances derivation of hematopoietic progenitor cells from human pluripotent stem cells. Stem Cell Res. 2017;21:32–9.
pubmed: 28368243 doi: 10.1016/j.scr.2017.03.014
Galat Y, Elcheva I, Dambaeva S, Katukurundage D, Beaman K, Iannaccone PM, et al. Application of small molecule CHIR99021 leads to the loss of hemangioblast progenitor and increased hematopoiesis of human pluripotent stem cells. Exp Hematol. 2018;65(38–48): e1.
Kim K, Abdal Dayem A, Gil M, Yang GM, Lee SB, Kwon OH, et al. 3,2’-Dihydroxyflavone improves the proliferation and survival of human pluripotent stem cells and their differentiation into hematopoietic progenitor cells. J Clin Med. 2020;9(3):669.
pubmed: 32131506 pmcid: 7141312 doi: 10.3390/jcm9030669
Smith BW, Rozelle SS, Leung A, Ubellacker J, Parks A, Nah SK, et al. The aryl hydrocarbon receptor directs hematopoietic progenitor cell expansion and differentiation. Blood. 2013;122(3):376–85.
pubmed: 23723449 pmcid: 3716202 doi: 10.1182/blood-2012-11-466722
Angelos MG, Kaufman DS. Advances in the role of the aryl hydrocarbon receptor to regulate early hematopoietic development. Curr Opin Hematol. 2018;25(4):273–8.
pubmed: 29697485 doi: 10.1097/MOH.0000000000000432
Mesquitta WT, Wandsnider M, Kang H, Thomson J, Moskvin O, Suknuntha K, et al. UM171 expands distinct types of myeloid and NK progenitors from human pluripotent stem cells. Sci Rep. 2019;9(1):6622.
pubmed: 31036928 pmcid: 6488662 doi: 10.1038/s41598-019-43054-4
Shim SH, Tufa D, Woods R, George TD, Shank T, Yingst A, et al. SAHA enhances differentiation of CD34
pubmed: 35349707 pmcid: 9154343 doi: 10.1093/stcltm/szac012
Suksamrarn A, Ponglikitmongkol M, Wongkrajang K, Chindaduang A, Kittidanairak S, Jankam A, et al. Diarylheptanoids, new phytoestrogens from the rhizomes of Curcuma comosa: isolation, chemical modification and estrogenic activity evaluation. Bioorg Med Chem. 2008;16(14):6891–902.
pubmed: 18554915 doi: 10.1016/j.bmc.2008.05.051
Winuthayanon W, Piyachaturawat P, Suksamrarn A, Ponglikitmongkol M, Arao Y, Hewitt SC, et al. Diarylheptanoid phytoestrogens isolated from the medicinal plant Curcuma comosa: biologic actions in vitro and in vivo indicate estrogen receptor-dependent mechanisms. Environ Health Perspect. 2009;117(7):1155–61.
pubmed: 19654927 pmcid: 2717144 doi: 10.1289/ehp.0900613
Tanhuad N, Thongsa-Ad U, Sutjarit N, Yoosabai P, Panvongsa W, Wongniam S, et al. Ex vivo expansion and functional activity preservation of adult hematopoietic stem cells by a diarylheptanoid from Curcuma comosa. Biomed Pharmacother. 2021;143: 112102.
pubmed: 34474347 doi: 10.1016/j.biopha.2021.112102
Bhukhai K, Fouquet G, Rittavee Y, Tanhuad N, Lakmuang C, Borwornpinyo S, et al. Enhancing erythropoiesis by a phytoestrogen diarylheptanoid from Curcuma comosa. Biomedicines. 2022;10(6):1427.
pubmed: 35740448 pmcid: 9219836 doi: 10.3390/biomedicines10061427
Tangprasittipap A, Jittorntrum B, Wongkummool W, Kitiyanant N, Tubsuwan A. Generation of induced pluripotent stem cells from peripheral blood CD34
pubmed: 28395748 doi: 10.1016/j.scr.2017.02.013
Daniel MG, Sachs D, Bernitz JM, Fstkchyan Y, Rapp K, Satija N, et al. Induction of human hemogenesis in adult fibroblasts by defined factors and hematopoietic coculture. FEBS Lett. 2019;593(23):3266–87.
pubmed: 31557312 pmcid: 6901732 doi: 10.1002/1873-3468.13621
Ponchio L, Duma L, Oliviero B, Gibelli N, Pedrazzoli P, Cuna GR. Mitomycin C as an alternative to irradiation to inhibit the feeder layer growth in long-term culture assays. Cytotherapy. 2000;2(4):281–6.
pubmed: 12042037 doi: 10.1080/146532400539215
Lange L, Hoffmann D, Schwarzer A, Ha TC, Philipp F, Lenz D, et al. Inducible forward programming of human pluripotent stem cells to hemato-endothelial progenitor cells with hematopoietic progenitor potential. Stem Cell Rep. 2020;15(1):274.
doi: 10.1016/j.stemcr.2020.05.019
Ronn RE, Guibentif C, Saxena S, Woods NB. Reactive oxygen species impair the function of CD90(
pubmed: 27641910 doi: 10.1002/stem.2503
Sutherland HJ, Lansdorp PM, Henkelman DH, Eaves AC, Eaves CJ. Functional characterization of individual human hematopoietic stem cells cultured at limiting dilution on supportive marrow stromal layers. Proc Natl Acad Sci USA. 1990;87(9):3584–8.
pubmed: 2333304 pmcid: 53946 doi: 10.1073/pnas.87.9.3584
Moore KA, Ema H, Lemischka IR. In vitro maintenance of highly purified, transplantable hematopoietic stem cells. Blood. 1997;89(12):4337–47.
pubmed: 9192756 doi: 10.1182/blood.V89.12.4337
Trakarnsanga K, Ferguson D, Daniels DE, Griffiths RE, Wilson MC, Mordue KE, et al. Vimentin expression is retained in erythroid cells differentiated from human iPSC and ESC and indicates dysregulation in these cells early in differentiation. Stem Cell Res Ther. 2019;10(1):130.
pubmed: 31036072 pmcid: 6489253 doi: 10.1186/s13287-019-1231-z
Tubsuwan A, Abed S, Deichmann A, Kardel MD, Bartholoma C, Cheung A, et al. Parallel assessment of globin lentiviral transfer in induced pluripotent stem cells and adult hematopoietic stem cells derived from the same transplanted beta-thalassemia patient. Stem Cells. 2013;31(9):1785–94.
pubmed: 23712774 doi: 10.1002/stem.1436
Mo JS, Park HW, Guan KL. The Hippo signaling pathway in stem cell biology and cancer. EMBO Rep. 2014;15(6):642–56.
pubmed: 24825474 pmcid: 4197875 doi: 10.15252/embr.201438638
Bucar S, Branco ADM, Mata MF, Milhano JC, Caramalho I, Cabral JMS, et al. Influence of the mesenchymal stromal cell source on the hematopoietic supportive capacity of umbilical cord blood-derived CD34(
pubmed: 34256848 pmcid: 8278708 doi: 10.1186/s13287-021-02474-8
McGrath Kathleen E, Frame Jenna M, Fegan Katherine H, Bowen James R, Conway Simon J, Catherman Seana C, et al. Distinct sources of hematopoietic progenitors emerge before HSCs and provide functional blood cells in the mammalian embryo. Cell Rep. 2015;11(12):1892–904.
pubmed: 26095363 pmcid: 4490098 doi: 10.1016/j.celrep.2015.05.036
Laurenti E, Gottgens B. From haematopoietic stem cells to complex differentiation landscapes. Nature. 2018;553(7689):418–26.
pubmed: 29364285 pmcid: 6555401 doi: 10.1038/nature25022
Niwa A, Heike T, Umeda K, Oshima K, Kato I, Sakai H, et al. A novel serum-free monolayer culture for orderly hematopoietic differentiation of human pluripotent cells via mesodermal progenitors. PLoS ONE. 2011;6(7): e22261.
pubmed: 21818303 pmcid: 3144871 doi: 10.1371/journal.pone.0022261
Tursky ML, Loi TH, Artuz CM, Alateeq S, Wolvetang EJ, Tao H, et al. Direct comparison of four hematopoietic differentiation methods from human induced pluripotent stem cells. Stem Cell Reports. 2020;15(3):735–48.
pubmed: 32763163 pmcid: 7486192 doi: 10.1016/j.stemcr.2020.07.009
Hansen M, Varga E, Aarts C, Wust T, Kuijpers T, von Lindern M, et al. Efficient production of erythroid, megakaryocytic and myeloid cells, using single cell-derived iPSC colony differentiation. Stem Cell Res. 2018;29:232–44.
pubmed: 29751281 doi: 10.1016/j.scr.2018.04.016
Salvagiotto G, Burton S, Daigh CA, Rajesh D, Slukvin II, Seay NJ. A defined, feeder-free, serum-free system to generate in vitro hematopoietic progenitors and differentiated blood cells from hESCs and hiPSCs. PLoS ONE. 2011;6(3): e17829.
pubmed: 21445267 pmcid: 3060827 doi: 10.1371/journal.pone.0017829
Ruiz JP, Chen G, Haro Mora JJ, Keyvanfar K, Liu C, Zou J, et al. Robust generation of erythroid and multilineage hematopoietic progenitors from human iPSCs using a scalable monolayer culture system. Stem Cell Res. 2019;41: 101600.
pubmed: 31710911 pmcid: 6953424 doi: 10.1016/j.scr.2019.101600
Tomellini E, Fares I, Lehnertz B, Chagraoui J, Mayotte N, MacRae T, et al. Integrin-alpha3 Is a functional marker of ex vivo expanded human long-term hematopoietic stem cells. Cell Rep. 2019;28(4):1063–73.
pubmed: 31340144 doi: 10.1016/j.celrep.2019.06.084
Bhukhai K, Suksen K, Bhummaphan N, Janjorn K, Thongon N, Tantikanlayaporn D, et al. A phytoestrogen diarylheptanoid mediates estrogen receptor/Akt/glycogen synthase kinase 3beta protein-dependent activation of the Wnt/beta-catenin signaling pathway. J Biol Chem. 2012;287(43):36168–78.
pubmed: 22936801 pmcid: 3476284 doi: 10.1074/jbc.M112.344747
Wu Y, Hirschi KK. Regulation of hemogenic endothelial cell development and function. Annu Rev Physiol. 2021;83:17–37.
pubmed: 33035429 doi: 10.1146/annurev-physiol-021119-034352
Gritz E, Hirschi KK. Specification and function of hemogenic endothelium during embryogenesis. Cell Mol Life Sci. 2016;73(8):1547–67.
pubmed: 26849156 pmcid: 4805691 doi: 10.1007/s00018-016-2134-0
Driskill JH, Pan D. Control of stem cell renewal and fate by YAP and TAZ. Nat Rev Mol Cell Biol. 2023;24(12):895–911.
pubmed: 37626124 doi: 10.1038/s41580-023-00644-5
Qin H, Hejna M, Liu Y, Percharde M, Wossidlo M, Blouin L, et al. YAP induces human naive pluripotency. Cell Rep. 2016;14(10):2301–12.
pubmed: 26947063 pmcid: 4807727 doi: 10.1016/j.celrep.2016.02.036
Wang S, Zhou L, Ling L, Meng X, Chu F, Zhang S, et al. The crosstalk between Hippo-YAP pathway and innate immunity. Front Immunol. 2020;11:323.
pubmed: 32174922 pmcid: 7056731 doi: 10.3389/fimmu.2020.00323
Damkham N, Issaragrisil S, Lorthongpanich C. Role of YAP as a mechanosensing molecule in stem cells and stem cell-derived hematopoietic cells. Int J Mol Sci. 2022;23(23):14634.
pubmed: 36498961 pmcid: 9737411 doi: 10.3390/ijms232314634
Laowtammathron C, Lorthongpanich C, Jiamvoraphong N, Srisook P, Klaihmon P, Kheolamai P, et al. Role of YAP in hematopoietic differentiation and erythroid lineage specification of human-induced pluripotent stem cells. Stem Cell Res Ther. 2023;14(1):279.
pubmed: 37775798 pmcid: 10543272 doi: 10.1186/s13287-023-03508-z
Howard A, Bojko J, Flynn B, Bowen S, Jungwirth U, Walko G. Targeting the Hippo/YAP/TAZ signalling pathway: novel opportunities for therapeutic interventions into skin cancers. Exp Dermatol. 2022;31(10):1477–99.
pubmed: 35913427 pmcid: 9804452 doi: 10.1111/exd.14655
Goode DK, Obier N, Vijayabaskar MS, Lie ALM, Lilly AJ, Hannah R, et al. Dynamic gene regulatory networks drive hematopoietic specification and differentiation. Dev Cell. 2016;36(5):572–87.
pubmed: 26923725 pmcid: 4780867 doi: 10.1016/j.devcel.2016.01.024
Kim KM, Mura-Meszaros A, Tollot M, Krishnan MS, Grundl M, Neubert L, et al. Taz protects hematopoietic stem cells from an aging-dependent decrease in PU.1 activity. Nat Commun. 2022;13(1):5187.
pubmed: 36057685 pmcid: 9440927 doi: 10.1038/s41467-022-32970-1
de la Grange P, Jolly A, Courageux C, Ben Brahim C, Leroy P. Genes coding for transcription factors involved in stem cell maintenance are repressed by TGF-beta and downstream of Slug/Snail2 in COPD bronchial epithelial progenitors. Mol Biol Rep. 2021;48(10):6729–38.
pubmed: 34436724 doi: 10.1007/s11033-021-06664-8
Liu X, Ye Y, Zhu L, Xiao X, Zhou B, Gu Y, et al. Niche stiffness sustains cancer stemness via TAZ and NANOG phase separation. Nat Commun. 2023;14(1):238.
pubmed: 36646707 pmcid: 9842735 doi: 10.1038/s41467-023-35856-y

Auteurs

Umnuaychoke Thongsa-Ad (U)

Department of Physiology, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand.

Anongnat Wongpan (A)

Department of Physiology, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand.

Wasinee Wongkummool (W)

Stem Cell Research Group, Institute of Molecular Biosciences, Mahidol University, Nakhon Pathom, 73170, Thailand.
Center of Multidisciplinary Technology for Advanced Medicine (CMUTEAM), Faculty of Medicine, Chiang Mai University, Chiang Mai, 50200, Thailand.

Phaewa Chaiwijit (P)

Department of Physiology, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand.

Kwanchanok Uppakara (K)

Chakri Naruebodindra Medical Institute, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Samut Prakan, 10540, Thailand.

Gorawin Chaiyakitpattana (G)

Department of Physiology, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand.

Passanan Singpant (P)

Stem Cell Research Group, Institute of Molecular Biosciences, Mahidol University, Nakhon Pathom, 73170, Thailand.

Pirut Tong-Ngam (P)

Stem Cell Research Group, Institute of Molecular Biosciences, Mahidol University, Nakhon Pathom, 73170, Thailand.

Amnat Chukhan (A)

Prima Scientific, 147/170-171 Baromrajchonnee, Arunamarin, Bangkok, 10700, Thailand.

Wachirachai Pabuprappap (W)

Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Ramkhamhaeng University, Bangkok, 10240, Thailand.

Sirapope Wongniam (S)

Center for Scientific Instrumentation and Platform Services Unit, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand.

Apichart Suksamrarn (A)

Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Ramkhamhaeng University, Bangkok, 10240, Thailand.

Suradej Hongeng (S)

Department of Pediatrics, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, 10400, Thailand.

Usanarat Anurathapan (U)

Department of Pediatrics, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, 10400, Thailand.

Kasem Kulkeaw (K)

Siriraj Integrative Center for Neglected Parasitic Diseases, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, 10700, Thailand.

Alisa Tubsuwan (A)

Stem Cell Research Group, Institute of Molecular Biosciences, Mahidol University, Nakhon Pathom, 73170, Thailand.

Kanit Bhukhai (K)

Department of Physiology, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand. kanit.bhu@mahidol.ac.th.

Classifications MeSH