California Institute for Regenerative Medicine awarded Sangamo $6.4M to develop its ZFP-genome editing technology for the treatment of beta-thalassemia (加州再生醫學研究所授予Sangamo公司了$ 6.4M開發其 ZFP 基因組編輯技術治療β-地中海貧血)

How the technology work?




Figure 1: Schematic to demonstrate the position and orientation of binding of a ZFNs pair to a specific DNA sequence to enable generation of a double strand break in the DNA.




Figure 2: Schematic to demonstrate potential outcomes of a double-strand break in DNA generated by a pair of ZFNs, gene disruption, gene correction or DNA insertion.


Original Source: http://www.sangamo.com/technology/zf-nucleases.html

Protein Injection Strategy

(Continuously Updating)

Paper 1: Wnt7a treatment ameliorates muscular dystrophy

Strategy :

1.  Wnt7a treatment efficiently induced satellite cell expansion and myofiber hypertrophy in treated mucles in mdx mice.
1. Wnt7a signaling through its receptor Fzd7 accelerates and augments regeneration by:
1. stimulating satellite stem cell expansion through the planar cell polarity pathway, as well as,
2. myofiber hypertrophy through the AKT/mammalian target of rapamycin (mTOR) anabolic pathway. 

Application:

1. Duchenne muscular dystrophy (DMD)

References: 

1. J. von Maltzahn, J.-M. Renaud, G. Parise, M. A. Rudnicki.Wnt7a treatment ameliorates muscular dystrophy.Proceedings of the National Academy of Sciences, 2012; DOI: 10.1073/pnas.1215765109

Cells Rejuvenation Strategy

(Continuously Updating)

Paper 1: Aged human cells rejuvenated by cytokine enhancement of biomaterials for surgical ventricular restoration.

Strategy : 

1.  cytokine enhancement of a biodegradable patch
1.  covalently immobilized 2 proangiogenic cytokines (vascular endothelial growth factor and basic fibroblast growth factor) onto porous collagen scaffolds.
2.  seeded human mesenchymal stromal cells from old donors into the scaffolds with  growth factors. 

Application: 

1. Surgical ventricular restoration (SVR)

References:

1. Kang K, Sun L, Xiao Y, Li SH, Wu J, Guo J, Jiang SL, Yang L, Yau TM, Weisel RD, Radisic M, Li RK. Aged human cells rejuvenated by cytokine enhancement of biomaterials for surgical ventricular restoration. J Am Coll Cardiol. 2012 Nov 20;60(21):2237-49. doi: 10.1016/j.jacc.2012.08.985. (Department of Cardiovascular Surgery, the Second Affiliated Hospital of Harbin Medical University, Harbin, China; Division of Cardiovascular Surgery and Toronto General Research Institute, University Health Network and Department of Surgery, Division of Cardiac Surgery, University of Toronto, Toronto, Ontario, Canada.) PMID: 23153846

Animal trials of Mesoblast's (ASX:MSB) mesenchymal stem cells in rheumatoid arthritis (RA) have encouraged the company press on with an FDA application for a phase II trial.

Original article:
http://www.lifescientist.com.au/article/431662/mesoblast_eyes_human_trials_stem_cell_treatment_rheumatoid_arthritis/


Mesoblast (ASX:MSB) is continuing to expand the applications for its off-the-shelf stem cell technology by pushing forward to conduct a phase II trial for rheumatoid arthritis (RA).

This follows encouraging results from preclinical trials of its adult stem cells in animal models of RA.

The company revealed that a single injection of its mesenchymal precursor cells (MPCs) was able to simultaneously inhibit multiple cytokines responsible for driving RA.

The MPCs concomitantly affected T cells, monocytes and synoviocytes to shut down TNF-alpha, IL-6 and IL-I7 cytokine pathways in sheep with collagen-induced arthritis.

Treatments that shut down any of these pathways alone are only moderately effective at treating RA, and often need to be administered chronically.

Based on the promising results, Mesoblast has arranged a meeting with the US FDA to discuss organising a phase II trial.

Subject to receiving approval, the company plans to conduct a randomised, placebo-controlled phase II trial in the fourth quarter.

During the pilot study, significant numbers of allogenic MPCs were detected in involved joints or lymph nodes of the arthritic sheep, but not in normal sheep. This suggests the MPCs selectively migrate to sites of immune-mediated inflammation.

Sheep were injected with three different quantities of MPCs. Joint tissue from sheep receiving the largest dose showed an 88% mean reduction in IL-6 levels, an 83% reduction in TNF-alpha levels and a 52% reduction in IL-I7 levels.

MPC-treated animals had a 31% mean reduction in histopathology severity scores compared to the control group.

Twenty autologous umbilical cord blood from private cord blood banks were evaluated for thaw characteristics. 研究對私人臍帶血庫的21個自體臍血進行解凍後的特點評估.

Twenty autologous umbilical cord blood from private cord blood banks were evaluated for thaw characteristics: "only 11% of evaluable autologous umbilical cord blood achieved the minimum total nucleated cell count of at least 9.0×10(8) to meet the National Cord Blood Inventory banking threshold and only 50% met the minimum of 5.0×10(8) TNC count for Food and Drug Administration cord blood licensure eligibility."

研究對私人臍帶血庫的21個自體臍血進行解凍後的特點評估“只有11％的自體臍血達到了9.0×10(8)的最低總核細胞數目, 以滿足國家臍帶血庫存的門檻; 此外, 亦只有50％的自體臍血達到了5.0×10(8)總核細胞數目,以滿足食品和藥物管理局臍帶血執照的資格.

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

http://www.ncbi.nlm.nih.gov/pubmed/22321210

Transfusion. 2012 Feb 10. doi: 10.1111/j.1537-2995.2011.03556.x. [Epub ahead of print]

Characteristics of thawed autologous umbilical cord blood.

Rosenau EH, Sugrue MW, Haller M, Fisk D, Kelly SS, Chang M, Hou W, Eldjerou L, Slayton W, Cogle CR, Wingard JR.

Source

From the Department of Medicine, the Department of Pediatrics, and the Department of Epidemiology and Health Policy Research, University of Florida, and Shands Hospital at the University of Florida, Gainesville, Florida; and the Department of Pediatrics Patient Care, University of Texas MD Anderson Cancer Center, Houston, Texas.

Abstract

BACKGROUND:

Autologous umbilical cord blood (AutoUCB) has historically been cryopreserved for potential use in hematopoietic transplantation. Increasingly, private AutoUCB banking is performed for therapies unavailable today. A Phase I trial using AutoUCB treatment for early pediatric Type 1 diabetes afforded us an opportunity to analyze characteristics of AutoUCBs.

STUDY DESIGN AND METHODS:

Twenty AutoUCBs from AABB-accredited private cord blood banks (CBBs) were evaluated for collection, processing, cryopreservation, and thaw characteristics. Using a standardized thaw-wash method, AutoUCBs were assessed for viable total nucleated cells (vTNCs), viable CD34+ (vCD34+), and colony-forming unit-granulocyte-macrophage counts. Postthaw %vTNC recoveries were compared against processing characteristics and analyzed according to processing method, cryopreservation volume, concentration, container, and length of storage.

RESULTS:

AutoUCB collection volumes (19.9-170 mL), cryopreserved TNC counts (7.6× 10(7) -3.34×10(9) ), %TNC processing recoveries (39%-100%), postthaw %vTNC recoveries (58%-100%), and %vCD34+ recoveries (26%-96%) varied widely. Regarding cell dose requirements, only 11% of evaluable AutoUCBs achieved the minimum TNC count of at least 9.0×10(8) to meet the National Cord Blood Inventory banking threshold, and only 50% met the minimum of 5.0×10(8) TNC count for Food and Drug Administration cord blood licensure eligibility. %vTNC recoveries correlated with %vCD34+ recoveries (R=0.7; p=0.03). Length of storage, cryopreservation volume, concentration, and container type did not affect postthaw %vTNC recoveries. CBB processing method, however, was associated with %vTNC postprocessing recoveries, with unmanipulated and plasma-depleted AutoUCBs having the highest postthaw %vTNC recovery, followed by RBC-depleted and density gradient-separated AutoUCBs.

CONCLUSION:

The high variability and low counts found in AutoUCB banking suggest that further standardization of characterization, collection, and processing procedures is needed.

© 2012 American Association of Blood Banks.

PMID:

 

22321210

 

[PubMed - as supplied by publisher]

A pilot study shows Intrabone Cord Blood Hematopoietic Stem Cell Transplantation is safe and feasible in pediatrics. 一項初步研究顯示:移植臍血造血幹細胞至骨髓內在兒科是安全和可行的.

http://www.ncbi.nlm.nih.gov/pubmed/22322938

J Pediatr Hematol Oncol. 2012 Feb 8. [Epub ahead of print]

Intrabone Cord Blood Hematopoietic Stem Cell Transplantation in a Subset of Very High-Risk Pediatric Patients: A Safety and Feasibility Pilot Study.

Saglio F, Berger M, Vassallo E, Nesi F, Gunetti M, Fazio L, Rustichelli D, Ferrero I, Fagioli F.

Source

*Pediatric Onco-Hematology, Stem Cell Transplantation and Cellular Therapy Division †Immunohematology and Transfusion Medicine Service, Regina Margherita Children's Hospital, Turin, Italy.

Abstract

The main limit of umbilical cord blood hematopoietic stem cell transplantation is a more difficult engraftment related to the number of cells infused per kilogram of recipient body weight. This limit makes the cord blood a suboptimal source of hematopoietic stem cells for transplantation in case of difficult engraftment situations. Direct intrabone cord blood (CB) injection has been recently investigated as a solution to cell dose problem in the adults population, but there is a lack of data concerning this approach in pediatric patients. Here, we describe 5 pediatric patients undergoing intrabone cord blood transplantation (IBCBT) for different diseases characterized by a high risk of posttransplant graft failure. The conditioning regimen differed according to the disease, whereas the GvHD prophylaxis consisted of cyclosporine, mycophenolate, and ATG. The median numbers of total nucleated cells infused and CD34 cells were 3.3×10/kg, 2×10/kg. All the patients showed complete hematological recovery and complete donor engraftment. No patient had secondary graft failure, whereas 1 patient relapsed 6 months after IBCBT. No patient died of transplant-related complications. Our results show that IBCBT is safe and feasible in pediatrics as well, and suggest that IBCBT might be an attractive option to overcome some limits of umbilical cord blood hematopoietic stem cell transplantation.

PMID:

 

22322938

 

[PubMed - as supplied by publisher]

Cord Blood "Stem Cell Educator therapy" may Reverse Type 1 Diabetes - Phase 1 / Phase 2 Clinical Study. 臍帶血“幹細胞教育療法”或逆轉1型糖尿病 (胰岛素依赖型糖尿病) - 階段1/ 階段2 臨床試驗.

Hypothesis: control autoimmune responses by altering regulatory T cells (Tregs) and human islet beta cell-specific T cell clones.

假設: 通過改變調節性T細胞(Treg細胞)和人體胰島β細胞特異性 T細胞克隆控制自身免疫反應”

------------------
Method: "Stem Cell Educator therapy": the diabetic patient's blood is circulated through a closed-loop system that separates lymphocytes (a class of immune cell that includes T cells) from the whole blood and co-cultures them with cord blood stem cells from healthy donors for two to three hours before returning the "re-educated lymphocytes" to the patient's circulation.

方法：“幹細胞教育療法”: 糖尿病患者的血液通過一個閉環系統, 分離血液中的淋巴細胞至臍帶血造血幹細胞的裝置, 培養兩到三個小時進行 “再教育淋巴細胞", 然後再回至病人身體 。

------------------
Results: the median daily dose of required insulin was down by 38% at week 12 for the six patients with moderate diabetes and by 25% for the patients with severe diabetes. There was no change in required insulin dose for the controls.

All the patients who had received the Stem Cell Educator therapy also showed improved levels of C-peptide, a biomarker used to measure how well beta cells are working (it is a protein fragment that is left behind when insulin is made in the pancreas).

結果: 實驗組中度和嚴重糖尿病每日所需胰島素劑量中位數下降分別38％和25％, 對照組所需的胰島素劑量沒有變化.

實驗組患者的C肽生物標誌水平也有相對提高.

-----------------
Clinical Study: Phase 1 / Phase 2 Study (15 patients), University of Illinois

臨床試驗: 階段1/ 階段2 研究(15名病人), 美國伊利諾伊大學

-------------------http://www.ncbi.nlm.nih.gov/pubmed?term=Reversal%20of%20type%201%20diabetes%20via%20islet%20%CE%B2%20cell%20regenerationfollowing%20immune%20modulation%20by%20cord%20blood-derivedmultipotent%20stem%20cells%20pubmed

http://www.medicalnewstoday.com/articles/240160.php

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

BMC Med. 2012 Jan 10;10(1):3. [Epub ahead of print]

Reversal of type 1 diabetes via islet beta cell regeneration following immune modulation by cord blood-derived multipotent stem cells.

Zhao Y, Jiang Z, Zhao T, Ye M, Hu C, Yin Z, Li H, Zhang Y, Diao Y, Li Y, Chen Y, Sun X, Fisk MB, Skidgel R, Holterman M, Prabhakar B, Mazzone T.

Abstract

ABSTRACT:

BACKGROUND:

Inability to control autoimmunity is the primary barrier to developing a cure for type 1 diabetes (T1D). Evidence that human cord blood-derived multipotent stem cells (CB-SCs) can control autoimmune responses by altering regulatory T cells (Tregs) and human islet beta cell-specific T cell clones offers promise for a new approach to overcome the autoimmunity underlying T1D.

METHODS:

We developed a procedure for Stem Cell Educator therapy in which a patient's blood is circulated through a closed-loop system that separate lymphocytes from the whole blood and briefly co-cultures them with adherent CB-SCs before returning them to the patient's circulation. In an open-label, phase1/phase 2 study, patients (n = 15) with T1D received one treatment with the Stem Cell Educator. Median age was 29 years (range, 15 to 41), and median diabetic history was 8 years (range, 1 to 21).

RESULTS:

Stem Cell Educator therapy was well tolerated in all participants with minimal pain from two venipunctures and no adverse events. Stem Cell Educator therapy can markedly improve C-peptide levels, reduce the median glycated hemoglobin A1C (HbA1C) values, and decrease the median daily dose of insulin in patients with some residual beta cell function (n = 6) and patients with no residual pancreatic islet beta cell function (n = 6). Treatment also produced an increase in basal and glucose-stimulated C-peptide levels through 40 weeks. However, participants in the Control Group (n = 3) did not exhibit significant change at any follow-up. Individuals who received Stem Cell Educator therapy exhibited increased expression of costimulating molecules (specifically, CD28 and ICOS), increases in the number of CD4+CD25+Foxp3+ Tregs, and restoration of Th1/Th2/Th3 cytokine balance.

CONCLUSIONS:

Stem Cell Educator therapy is safe, and in individuals with moderate or severe T1D, a single treatment produces lasting improvement in metabolic control. Initial results indicate Stem Cell Educator therapy reverses autoimmunity and promotes regeneration of islet beta cells. Successful immune modulation by CB-SCs and the resulting clinical improvement in patient status may have important implications for other autoimmune and inflammation-related diseases without the safety and ethical concerns associated with conventional stem cell-based approaches. 

Trial registration: ClinicalTrials.gov number, NCT01350219.

PMID:

 

22233865

 

[PubMed - as supplied by publisher] 

Free full text