Fetal liver CD34+ contain human immune and endothelial progenitors and mediate solid tumor rejection in NOG mice
Background: Transplantation of CD34+ hematopoietic stem and progenitor cells (HSPC) into immunodeficient mice is an established method to generate humanized mice harbouring a human immune system. Different sources and methods for CD34+ isolation have been employed by various research groups, resulti...
| Main Authors: | , , , , , , , , , , , , |
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| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
| Published: |
2024
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| Subjects: | |
| Online Access: | https://hdl.handle.net/10356/179603 |
| _version_ | 1826118941389357056 |
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| author | Celhar, Teja Li, Xinyi Zhao, Yunqian Tay, Hui Chien Lee, Andrea Liew, Hui Hua Shepherdson, Edwin Kunxiang Rajarethinam, Ravisankar Fan, Yiping Mak, Anselm Chan, Jerry Kok Yen Singhal, Amit Takahashi, Takeshi |
| author2 | Lee Kong Chian School of Medicine (LKCMedicine) |
| author_facet | Lee Kong Chian School of Medicine (LKCMedicine) Celhar, Teja Li, Xinyi Zhao, Yunqian Tay, Hui Chien Lee, Andrea Liew, Hui Hua Shepherdson, Edwin Kunxiang Rajarethinam, Ravisankar Fan, Yiping Mak, Anselm Chan, Jerry Kok Yen Singhal, Amit Takahashi, Takeshi |
| author_sort | Celhar, Teja |
| collection | NTU |
| description | Background: Transplantation of CD34+ hematopoietic stem and progenitor cells (HSPC) into immunodeficient mice is an established method to generate humanized mice harbouring a human immune system. Different sources and methods for CD34+ isolation have been employed by various research groups, resulting in customized models that are difficult to compare. A more detailed characterization of CD34+ isolates is needed for a better understanding of engraftable hematopoietic and potentially non-hematopoietic cells. Here we have performed a direct comparison of CD34+ isolated from cord blood (CB-CD34+) or fetal liver (FL-CD34+ and FL-CD34+CD14−) and their engraftment into immunocompromised NOD/Shi-scid Il2rgnull (NOG) mice. Methods: NOG mice were transplanted with either CB-CD34+, FL-CD34+ or FL-CD34+CD14− to generate CB-NOG, FL-NOG and FL-CD14−-NOG, respectively. After 15–20 weeks, the mice were sacrificed and human immune cell reconstitution was assessed in blood and several organs. Liver sections were pathologically assessed upon Haematoxylin and Eosin staining. To assess the capability of allogenic tumor rejection in CB- vs. FL-reconstituted mice, animals were subcutaneously engrafted with an HLA-mismatched melanoma cell line. Tumor growth was assessed by calliper measurements and a Luminex-based assay was used to compare the cytokine/chemokine profiles. Results: We show that CB-CD34+ are a uniform population of HSPC that reconstitute NOG mice more rapidly than FL-CD34+ due to faster B cell development. However, upon long-term engraftment, FL-NOG display increased numbers of neutrophils, dendritic cells and macrophages in multiple tissues. In addition to HSPC, FL-CD34+ isolates contain non-hematopoietic CD14+ endothelial cells that enhance the engraftment of the human immune system in FL-NOG mice. We demonstrate that these CD14+CD34+ cells are capable of reconstituting Factor VIII-producing liver sinusoidal endothelial cells (LSEC) in FL-NOG. However, CD14+CD34+ also contribute to hepatic sinusoidal dilatation and immune cell infiltration, which may culminate in a graft-versus-host disease (GVHD) pathology upon long-term engraftment. Finally, using an HLA-A mismatched CDX melanoma model, we show that FL-NOG, but not CB-NOG, can mount a graft-versus-tumor (GVT) response resulting in tumor rejection. Conclusion: Our results highlight important phenotypical and functional differences between CB- and FL-NOG and reveal FL-NOG as a potential model to study hepatic sinusoidal dilatation and mechanisms of GVT. |
| first_indexed | 2024-10-01T04:51:49Z |
| format | Journal Article |
| id | ntu-10356/179603 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T04:51:49Z |
| publishDate | 2024 |
| record_format | dspace |
| spelling | ntu-10356/1796032024-08-18T15:39:36Z Fetal liver CD34+ contain human immune and endothelial progenitors and mediate solid tumor rejection in NOG mice Celhar, Teja Li, Xinyi Zhao, Yunqian Tay, Hui Chien Lee, Andrea Liew, Hui Hua Shepherdson, Edwin Kunxiang Rajarethinam, Ravisankar Fan, Yiping Mak, Anselm Chan, Jerry Kok Yen Singhal, Amit Takahashi, Takeshi Lee Kong Chian School of Medicine (LKCMedicine) Singapore Immunology Network, A*STAR A*STAR Infectious Diseases Labs Medicine, Health and Life Sciences Humanized mice Fetal liver CD34 Background: Transplantation of CD34+ hematopoietic stem and progenitor cells (HSPC) into immunodeficient mice is an established method to generate humanized mice harbouring a human immune system. Different sources and methods for CD34+ isolation have been employed by various research groups, resulting in customized models that are difficult to compare. A more detailed characterization of CD34+ isolates is needed for a better understanding of engraftable hematopoietic and potentially non-hematopoietic cells. Here we have performed a direct comparison of CD34+ isolated from cord blood (CB-CD34+) or fetal liver (FL-CD34+ and FL-CD34+CD14−) and their engraftment into immunocompromised NOD/Shi-scid Il2rgnull (NOG) mice. Methods: NOG mice were transplanted with either CB-CD34+, FL-CD34+ or FL-CD34+CD14− to generate CB-NOG, FL-NOG and FL-CD14−-NOG, respectively. After 15–20 weeks, the mice were sacrificed and human immune cell reconstitution was assessed in blood and several organs. Liver sections were pathologically assessed upon Haematoxylin and Eosin staining. To assess the capability of allogenic tumor rejection in CB- vs. FL-reconstituted mice, animals were subcutaneously engrafted with an HLA-mismatched melanoma cell line. Tumor growth was assessed by calliper measurements and a Luminex-based assay was used to compare the cytokine/chemokine profiles. Results: We show that CB-CD34+ are a uniform population of HSPC that reconstitute NOG mice more rapidly than FL-CD34+ due to faster B cell development. However, upon long-term engraftment, FL-NOG display increased numbers of neutrophils, dendritic cells and macrophages in multiple tissues. In addition to HSPC, FL-CD34+ isolates contain non-hematopoietic CD14+ endothelial cells that enhance the engraftment of the human immune system in FL-NOG mice. We demonstrate that these CD14+CD34+ cells are capable of reconstituting Factor VIII-producing liver sinusoidal endothelial cells (LSEC) in FL-NOG. However, CD14+CD34+ also contribute to hepatic sinusoidal dilatation and immune cell infiltration, which may culminate in a graft-versus-host disease (GVHD) pathology upon long-term engraftment. Finally, using an HLA-A mismatched CDX melanoma model, we show that FL-NOG, but not CB-NOG, can mount a graft-versus-tumor (GVT) response resulting in tumor rejection. Conclusion: Our results highlight important phenotypical and functional differences between CB- and FL-NOG and reveal FL-NOG as a potential model to study hepatic sinusoidal dilatation and mechanisms of GVT. Agency for Science, Technology and Research (A*STAR) National Research Foundation (NRF) Published version The CIEA-KKH-SIgN Humanized NOG Mice for Immuno Therapy (HuNIT) platform is supported by the Industry Alignment Fund – Pre-positioning Programme (IAF-PP; #H16/01/a0/002). JKYC is supported by Singapore’s Ministry of Health (MOH-001221-01, MOH-000932-00) and National Medical Research Council (CIRG/1484/2018, NMRC CSA (SI)/008/2016, NMRC STaR22jul-0004). The Immunomonitoring platform is supported by the National Research Foundation (NRF), Immunomonitoring Service Platform (ISP) grant (Ref: NRF2017_SISFP09). 2024-08-13T01:14:52Z 2024-08-13T01:14:52Z 2024 Journal Article Celhar, T., Li, X., Zhao, Y., Tay, H. C., Lee, A., Liew, H. H., Shepherdson, E. K., Rajarethinam, R., Fan, Y., Mak, A., Chan, J. K. Y., Singhal, A. & Takahashi, T. (2024). Fetal liver CD34+ contain human immune and endothelial progenitors and mediate solid tumor rejection in NOG mice. Stem Cell Research and Therapy, 15(1). https://dx.doi.org/10.1186/s13287-024-03756-7 1757-6512 https://hdl.handle.net/10356/179603 10.1186/s13287-024-03756-7 15 2-s2.0-85195624408 1 15 en H16/01/a0/002 NRF2017_SISFP09 Stem Cell Research and Therapy © The Author(s) 2024. Open Access. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. application/pdf |
| spellingShingle | Medicine, Health and Life Sciences Humanized mice Fetal liver CD34 Celhar, Teja Li, Xinyi Zhao, Yunqian Tay, Hui Chien Lee, Andrea Liew, Hui Hua Shepherdson, Edwin Kunxiang Rajarethinam, Ravisankar Fan, Yiping Mak, Anselm Chan, Jerry Kok Yen Singhal, Amit Takahashi, Takeshi Fetal liver CD34+ contain human immune and endothelial progenitors and mediate solid tumor rejection in NOG mice |
| title | Fetal liver CD34+ contain human immune and endothelial progenitors and mediate solid tumor rejection in NOG mice |
| title_full | Fetal liver CD34+ contain human immune and endothelial progenitors and mediate solid tumor rejection in NOG mice |
| title_fullStr | Fetal liver CD34+ contain human immune and endothelial progenitors and mediate solid tumor rejection in NOG mice |
| title_full_unstemmed | Fetal liver CD34+ contain human immune and endothelial progenitors and mediate solid tumor rejection in NOG mice |
| title_short | Fetal liver CD34+ contain human immune and endothelial progenitors and mediate solid tumor rejection in NOG mice |
| title_sort | fetal liver cd34 contain human immune and endothelial progenitors and mediate solid tumor rejection in nog mice |
| topic | Medicine, Health and Life Sciences Humanized mice Fetal liver CD34 |
| url | https://hdl.handle.net/10356/179603 |
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