Extended Survival of Glioblastoma Patients After Chemoprotective HSC Gene Therapy

• stm.sciencemag.org
• Jennifer E. Adair
• May 9th, 2012
• view original

Abstract

Chemotherapy with alkylating agents for treating malignant disease results in myelosuppression that can significantly limit dose escalation and potential clinical efficacy. Gene therapy using mutant methylguanine methyltransferase (P140K) gene–modified hematopoietic stem and progenitor cells may circumvent this problem by abrogating the toxic effects of chemotherapy on hematopoietic cells. However, this approach has not been evaluated clinically. Here, we show efficient polyclonal engraftment of autologous P140K-modified hematopoietic stem and progenitor cells in three patients with glioblastoma. Increases in P140K-modified cells after transplant indicate selection of gene-modified hematopoietic repopulating cells. Longitudinal retroviral integration site (RIS) analysis identified more than 12,000 unique RISs in the three glioblastoma patients, with multiple clones present in the peripheral blood of each patient throughout multiple chemotherapy cycles. To assess safety, we monitored RIS distribution over the course of chemotherapy treatments. Two patients exhibited emergence of prominent clones harboring RISs associated with the intronic coding region of PRDM16 (PR domain–containing 16) or the 3′ untranslated region of HMGA2 (high-mobility group A2) genes with no adverse clinical outcomes. All three patients surpassed the median survival for glioblastoma patients with poor prognosis, with one patient alive and progression-free more than 2 years after diagnosis. Thus, transplanted P140K-expressing hematopoietic stem and progenitor cells are chemoprotective, potentially maximizing the drug dose that can be administered.

#Cordblood #Stemcell derived DCs Generate Potent Antigen-Specific Immune Responses and Anti-tumor Effects.

Cord Blood Stem Cell-derived DCs Generate Potent Antigen-Specific Immune Responses and Anti-tumor Effects

http://www.clinsci.org/cs/imps/abs/CS20110272.htm?elq=927654c999f649559560fde6226abd79

Clinical Science (2012) Immediate Publication, doi:10.1042/CS20110272

Full Text PDF

Citing articles

Cord Blood Stem Cell-derived DCs Generate Potent Antigen-Specific Immune Responses and Anti-tumor Effects

Ming-Cheng Chang, Chien-Nan Lee, Yu-Li Chen, Ying-Cheng Chiang, Wei-Zen Sun, Yu-Hao Hu, CHi-An Chen and Wen-Fang Cheng

National Taiwan University Hospital, Taipei, Taiwan. wenfangcheng@yahoo.com

---

This study aims to evaluate if cord blood stem cells (CBSCs) can be new source of dendritic cells (DCs) which can generate more potent antigen-specific immune responses and anti-tumor effects. The CBSCs and peripheral blood mononuclear cells (PBMCs) were collected, cultured and differentiated into DCs. Surface markers, secreting cytokines, antigen presentation activity, antigen-specific cell-mediated immunity and cytotoxic killing effects induced by these two origins of DCs were evaluated and compared. The CBSCs expanded for ~17-fold by ex vivo culture. The expressions of surface markers in CBSC-derived DCs were higher than those in PBMC-derived DCs treated with LPS. The CBSC-derived DCs mainly secreted IL-6, IL-10, and TNF-a, while PBMC-derived DCs mainly secreted IL-5 and IFN-γ. The CBSC-derived DCs had better antigen presentation abilities when stimulated with LPS or TNF-a, induced higher numbers of IFN-g-secreting, antigen-specific CD8⁺ T lymphocytes by ELLIspot assay, and stimulated stronger antigen-specific CTL activities (p<0.01, one-way ANOVA). The CBSC-derived DCs showed quicker and stronger ERK and Akt phosphorylation, and weaker p38 phosphorylation than PBMC-derived DCs when stimulated with LPS. The CBSC-derived DCs have abilities of inducing stronger antigen-specific immunity and more potent anti-tumor effects. The CBSCs can be a good source of DCs in the strategy of DC-based cancer vaccine and immunotherapy.

---

doi:10.1042/CS20110272 Received 25 May 2011/12 January 2012; Accepted 23 January 2012 Published as Immediate Publication 23 January 2012

Stem Cells Dev. 2011 Aug;20(8):1371-81. Epub 2011 Jan 31.

Mesenchymal stem cells secreting angiopoietin-like-5 support efficient expansion of human hematopoietic stem cells without compromising their repopulating potential.

Khoury M, Drake A, Chen Q, Dong D, Leskov I, Fragoso MF, Li Y, Iliopoulou BP, Hwang W, Lodish HF, Chen J.

Source

Department of Biology, Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.

Abstract

Clinical and preclinical applications of human hematopoietic stem cells (HSCs) are often limited by scarcity of cells. Expanding human HSCs to increase their numbers while maintaining their stem cell properties has therefore become an important area of research. Here, we report a robust HSC coculture system wherein cord blood CD34(+) CD133(+) cells were cocultured with mesenchymal stem cells engineered to express angiopoietin-like-5 in a defined medium. After 11 days of culture, SCID repopulating cells were expanded ~60-fold by limiting dilution assay in NOD-scid Il2rg(-/-) (NSG) mice. The cultured CD34(+) CD133(+) cells had similar engraftment potential to uncultured CD34(+) CD133(+) cells in competitive repopulation assays and were capable of efficient secondary reconstitution. Further, the expanded cells supported a robust multilineage reconstitution of human blood cells in NSG recipient mice, including a more efficient T-cell reconstitution. These results demonstrate that the expanded CD34(+) CD133(+) cells maintain both short-term and long-term HSC activities. To our knowledge, this ~60-fold expansion of SCID repopulating cells is the best expansion of human HSCs reported to date. Further development of this coculture method for expanding human HSCs for clinical and preclinical applications is therefore warranted.

PMID:

 

21142526

 

[PubMed - indexed for MEDLINE] 

PMCID: PMC3148832

 [Available on 2012/8/1]

Int J Lab Hematol. 2011 Oct;33(5):516-25. doi: 10.1111/j.1751-553X.2011.01324.x. Epub 2011 Apr 14.

Ex vivo amplification of human hematopoietic stem and progenitor cells in an alginate three-dimensional culture system.

Yuan Y, Tse KT, Sin FW, Xue B, Fan HH, Xie Y, Xie Y.

Source

Section of Marine Ecology and Biotechnology, Division of Life Science, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong SAR, China.

Abstract

INTRODUCTION:

One of the key factors critical for a successful human cord blood transplantation in treating patients with hematopoietic disorders is the number of hematopoietic stem/progenitor cells derived from human cord blood. Here, we report an alginate three-dimensional (3D) culture system for the expansion of CD34+ cells in cord blood mononuclear cells (CBMCs).

METHODS:

Cord blood mononuclear cells were isolated from human cord blood and encapsulated in 3D alginate beads. The cells were grown with different concentrations of cytokines. At day 0, 3, 6, 9, and 12, respectively, the percentage of CD34+ cells was quantified by flow cytometry. Colony-forming cell assay was performed to determine the potential of hematopoietic reconstruction of the amplified cells under the 3D culture system.

RESULTS:

After culturing for 12 days, the CBMCs encapsulated in the 3D alginate beads were amplified 5.89 ± 0.72 fold, CD34+ cells increased from 2.60 ± 0.52% to 13.27 ± 2.65%, and the colony-forming assay showed that the colony-forming unit-granulocyte/granulocyte-macrophage (CFU-G/GM) increased from 363.34 ± 34.47/10⁵ cells to 3423.33 ± 645.14/10⁵ cells (P < 0.001). In comparison, the conventional two-dimensional (2D) culture system showed that the CBMCs, CD34+ cells and the CFU-G/GM were 0.68 ± 0.16 fold, 0.45 ± 0.17%, and 532.92 ± 82.97/10⁵ cells, respectively.

CONCLUSION:

This study demonstrates a new and efficient method to amplify the CD34+ human cord blood hematopoietic stem/progenitor cells in a 3D alginate culture system ex vivo for extended periods while retaining the hematopoietic reconstruction capacity.

© 2011 Blackwell Publishing Ltd.

PMID:

 

21492411

 

[PubMed - indexed for MEDLINE]

Combined growth factors enhanced angiogenic potential of #cordblood derived mononuclear cells transplanted to ischemic limbs.

http://www.ncbi.nlm.nih.gov/pubmed/21782826

Potential novel therapy for the treatment of ischemic cardiovascular disease due to the cultured cord blood MNCs have increased level of engraftment and angiogenic potential.

---------------------

J Mol Cell Cardiol. 2011 Nov;51(5):702-12. Epub 2011 Jul 19.

Combined growth factors enhanced angiogenic potential of cord blood-derived mononuclear cells transplanted to ischemic limbs.

Kim MH, Zhang HZ, Kim SW.

Source

Department of Cardiology, College of Medicine, Dong-A University, Regional Clinical Center, Dong-A University Hospital, Busan, South Korea.

Abstract

Stem cell therapy has recently been limited by poor engraftment and the marginal influence of the administered cells. This study aimed to enhance the survival and angiogenic capacity of human umbilical cord blood (UCB)-derived mononuclear cells (MNCs) and to demonstrate their therapeutic effects on experimental ischemia. A specific culture medium containing five growth factors (Flt-3L, EGF, TPO, FGF and IGF-1) augmented cell proliferation, adhesion potential as well as stimulated MNCs to become progenitor-like cells. In addition, qRT-PCR demonstrated that MNCs cultured with these five growth factors (5f-MNCs) markedly up-regulated multiple angiogenic, arteriogenic and anti-apoptotic factors compared with uncultured MNCs. In an ischemic hindlimb model, the injection of 5f-MNCs prevented limb loss and augmented blood perfusion, capillary density, vascular maturation and angiogenic cytokines in the affected tissues. In addition, the 5f-MNCs exhibited an increased engraftment rate and an endothelial phenotype and stimulated angiogenic factors in ischemic hindlimbs as demonstrated by flow cytometric, immunohistochemical and qRT-PCR analyses. Taken together, these data suggest that 5f-MNCs could be used as a novel therapy for the treatment of ischemic cardiovascular disease due to their increased level of engraftment and angiogenic potential.

Copyright © 2011 Elsevier Ltd. All rights reserved.

PMID:

 

21782826

 

[PubMed - indexed for MEDLINE]

骨髓中的內皮細胞和血管周圍細胞是負責造血幹細胞的維持功能.Study demonstrates Endothelial and perivascular cells are responsible for Blood #stemcell maintenance

造血幹細胞壁龕研究: 小鼠體內實驗: 運用選擇性敲入綠色熒光的模仿基因, 成功演示骨髓中的內皮細胞( Endothelial cells)和血管周圍細胞 (perivascular cells)是負責 造血幹細胞 (HSC)的維持功能.


Conditional knock-in of a loss-of-function SCF-GFP gene demonstrates Endothelial and perivascular cells are responsible for HSC maintenance.


http://www.nature.com/nature/journal/v481/n7382/full/nature10783.html


----------------

Nature. 2012 Jan 25;481(7382):457-62. doi: 10.1038/nature10783.

Endothelial and perivascular cells maintain haematopoietic stem cells.

Ding L, Saunders TL, Enikolopov G, Morrison SJ.

Source

Howard Hughes Medical Institute, Department of Pediatrics, University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA.

Abstract

Several cell types have been proposed to create niches for haematopoietic stem cells (HSCs). However, the expression patterns of HSC maintenancefactors have not been systematically studied and no such factor has been conditionally deleted from any candidate niche cell. Thus, the cellular sources of these factors are undetermined. Stem cell factor (SCF; also known as KITL) is a key niche component that maintains HSCs. Here, using Scf(gfp) knock-in mice, we found that Scf was primarily expressed by perivascular cells throughout the bone marrow. HSC frequency and function were not affected when Scf was conditionally deleted from haematopoietic cells, osteoblasts, nestin-cre- or nestin-creER-expressing cells. However, HSCs were depleted from bone marrow when Scf was deleted from endothelial cells or leptin receptor (Lepr)-expressing perivascular stromal cells. Most HSCs were lost when Scf was deleted from both endothelial and Lepr-expressing perivascular cells. Thus, HSCs reside in a perivascular niche in which multiple cell types express factors that promote HSC maintenance.

Comment in

• Nature. 2012;481(7382):453-5.

一個不需要預輻射照處理的免疫缺陷小鼠模型: Engraftment of human HSC in non-irradiated newborn NOD-scid IL2rγnull mice is enhanced by transgenic expression of membrane-bound human SCF.

一個新的使用免疫缺陷小鼠模型(immunodeficient mouse model)的方法: 不需要預輻射照處理 (pre-irradiation treatment)也可以移植入人類臍帶血造血幹細胞(Cord Blood HSC), 供應人類造血和免疫系統模型作臨床前研究(translational research).

------------------------

Blood. 2012 Jan 12. [Epub ahead of print]

Engraftment of human HSC in non-irradiated newborn NOD-scid IL2rγnull mice is enhanced by transgenic expression of membrane-bound human SCF.

Brehm MA, Racki WJ, Leif J, Burzenski L, Hosur V, Wetmore A, Gott B, Herlihy M, Ignotz R, Dunn R, Shultz LD, Greiner DL.

Source

Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA, United States;

Abstract

Immunodeficient mice engrafted with human hematopoietic stem cells (HSC) support multi-disciplinary translational experimentation, including the study of human hematopoiesis. Heightened levels of human HSC engraftment are observed in immunodeficient mice expressing mutations in the IL2-receptor common gamma chain (IL2rg) gene, including NOD-scid IL2rγ(null) (NSG) mice. Engraftment of human HSC requires pre-conditioning of immunodeficient recipients, usually with irradiation. Such preconditioning increases the expression of stem cell factor (SCF), which is critical for HSC engraftment, proliferation and survival. We hypothesized that transgenic expression of human membrane-bound stem cell factor Tg(hu-mSCF)] would increase levels of human HSC engraftment in non-irradiated NSG mice and eliminate complications associated with irradiation. Surprisingly, detectable levels of human CD45+ cell chimerism were observed following transplantation of cord blood-derived human HSC into non-irradiated adult as well as newborn NSG mice. However, transgenic expression of human mSCF enabled heightened levels of human hematopoietic cell chimerism in the absence of irradiation. Moreover, non-irradiated NSG-Tg(hu-mSCF) mice engrafted as newborns with human HSC rejected human skin grafts from a histoincompatible donor, indicating the development of a functional human immune system. These data provide a new immunodeficient mouse model that does not require irradiation pre-conditioning for human HSC engraftment and immune system development.

PMID:

 

22246028

 

[PubMed - as supplied by publisher]