Interview with Masayo Takahashi of RIKEN re: iPS Trial
RIKEN to resume retinal iPS transplant
Speaking with Masayo Takahashi: Targeting age-related macular degeneration in the first iPSC clinical trial
In this interview, Masayo Takahashi, Project Leader for Laboratory for Retinal Regeneration, RIKEN Center for Developmental Biology (Kobe, Japan) explains the techniques being utilized in the first induced pluripotent stem cell (iPSC) clinical trial in humans. Dr Takahashi also reveals how they may change when the trial restarts, having been halted in 2015.
Masayo Takahashi (MD, PhD)
Masayo Takahashi MD, PhD is Project Leader of the Laboratory for Retinal Regeneration at RIKEN (Kobe, Japan). She received her MD from Kyoto University (Kyoto, Japan) in 1986, and her PhD in Medicine at the same institution in 1992. After serving as an assistant professor in the Department of Ophthalmology, Kyoto University Hospital, she moved to the Salk Institute (CA, USA) in 1995, where she first learned of the potential of stem cells as a tool for retinal therapy. She returned to the same hospital in 1997, and since 2001 has served as an associate professor at the Translational Research Center (Kyoto, Japan). She joined RIKEN as a team leader of the retinal regeneration team in 2006. Her team started a pilot clinical study of autologous iPS cell-derived RPE cell sheets for exudative aged-related macular degeneration in 2013. The first RPE cell sheet graft was transplanted in September 2014.
Why did you choose age-related macular degeneration as the target for the iPSC therapy?
My research using stem cells began in 1995 and I have been trying to develop a therapy ever since. I researched embryonic stem cells (ESCs) and finally came to study iPSCs. From iPSCs, we can develop different retinal cells, including retinal pigment epithelial (RPE) cells and photoreceptor cells, and we are currently working on a therapy using both of these. However, photoreceptor cells are neurons so it can be difficult to make them work in the body, whereas RPE cells are not so it is easier to develop treatment. We found that RPE cells were most effective in treating age-related macular degeneration, which afflicts many people, so this is the target we chose first. [Ed: A study published in the Lancet in 2014 estimated that 196 million people will suffer from either dry- or wet-type age-related macular degeneration, rising to 288 million in 20401.]
What challenges have you faced in planning and executing the trial?
It’s very difficult to make and maintain good iPSCs, but we have very talented technicians who can make good iPSCs and culture them properly. In the first trial, we made 30 lines of iPSCs from one patient then selected the good iPSCs. The purity of the end product is also very important; there should be no bad cell contamination.
What techniques did you utilize to select the good iPSCs?
We chose the best cell lines by looking at their morphology first. Our technicians are very experienced so by looking at characteristics such as the colors, the smoothness of the cell surface and the dividing speed, they can tell which cells are good and bad. We then confirmed that the cells were good using gene analysis.
In 2015, the trial was halted. What happened, and will any changes be made to the trial design when it starts up again?
The first clinical research was halted because the law for regenerative medicine in Japan changed, but we’re now preparing the next clinical study. We will use three different protocols including allogeneic iPSCs. In the first trial, we used only autologous iPSCs, but next time we will use both. We won’t use immunosuppressive drugs, which is a big challenge when using allogeneic cells, but we will utilize human leukocyte antigen (HLA) typing to minimize any problems this may cause.
What does the future hold for iPSC therapy?
I think it’s very promising. Without a doubt, regenerative medicine will become a big field in the future. In fact, preventative and regenerative medicine will be two big fields, and iPSCs and ESCs have the potential to industrialize the cell therapy field. iPSCs and ESCs can be expanded so we can make many cells for many patients and really build regenerative medicine as an industry.
- • Research fund from Healios K.K. and Sumitomo Dainippon Pharma Co., Ltd.
- Wong WL, Su X, Li X. Global prevalence of age-related macular degeneration and disease burden projection for 2020 and 2040: a systematic review and meta-analysis. The Lancet Global Health. 2(2), e106 - e116.
- Update article on approval of Allo iPSC cells for human application
RIKEN to resume retinal iPS transplant
KOBE (Jiji Press) — The government-affiliated research institute RIKEN said Monday that it will resume its clinical study in which retinal tissues developed from iPS cells will be transplanted in an eye disease patient, in cooperation with Kyoto University and other medical institutes.
In 2014, the RIKEN Center for Developmental Biology, or CDB, successfully conducted a retinal transplant using induced pluripotent stem cells for the first time in the world. But its second trial was suspended due to a gene abnormality found in iPS cells.
In the first trial, iPS cells were created from cells taken from the patient who underwent the transplant.
Next time, the study team, led by Masayo Takahashi, project leader at the CDB, plans to use iPS cells created from mature cells of some others, since the first operation proved using a patient’s own cells is time-consuming and costly.
For the second trial, the CDB will develop retinal tissues from iPS cells supplied by Kyoto University’s Center for iPS Cell Research and Application, headed by Nobel laureate Shinya Yamanaka, the creator of the pluripotent cells.
Transplants of CDB-developed retinal tissues will be conducted at Kobe City Medical Center General Hospital and Osaka University Hospital.
The four institutes have signed an agreement to strengthen their collaboration. The partnership is “encouraging,” Takahashi said at a press conference on Monday. She expressed eagerness to carry out the next operation early next year.
Yamanaka said at the same news conference the he was heartened by the four institutes teaming up to push the study forward.
Credits: The Japan News by The Yomiuri Shimbun / Jiji Press
The Quest for Advanced Regenerative Medicine
Foreign Firms Drawn by Japan’s Expedited Approval System
Foreign Firms Drawn by Japan’s Expedited Approval System
Regenerative medicine is drawing attention as a form of treatment using people’s cells and a method that can be used for testing new pharmaceuticals. Japan has been leading the world in this field, as exemplified by the research into induced pluripotent stem cells (iPS cells) by Dr. Shinya Yamanaka, winner of the 2012 Nobel Prize in Physiology or Medicine. The global market for regenerative medicine is seen as growing to a huge JPY 38 trillion (USD 345 billion) by 2050. In Japan, government and industry are working hand in hand to advance this field with measures including bold regulatory reform and active promotion of investment by foreign corporations.
In November 2014 Japan became the first country in the world to adopt an expedited approval system for regenerative medical products and to allow outsourced cell culturing. Two products were approved under the new system within a year of its adoption. Thanks to this system, it has become possible in some cases to secure approval of new regenerative medical products in Japan prior to their approval in other countries.
In 2015 three government ministries—the Ministry of Education, Culture, Sports, Science and Technology; the Ministry of Health, Labour and Welfare; and the Ministry of Economy, Trade and Industry—joined in establishing the Japan Agency for Medical Research and Development. Through this agency the three ministries have created a unified setup for promotion of research and development. Meanwhile, the Japan External Trade Organization (JETRO) has taken the lead in efforts to support foreign companies that invest in regenerative medicine in Japan, with some JPY 1 billion (USD 9 million) in financial support available. Thanks to such initiatives, foreign companies have come to see Japan’s regenerative medicine market as offering opportunities.
Japanese industry is also welcoming the entry of foreign participants. As early as 2011, companies involved in this field joined in launching the Forum for Innovative Regenerative Medicine (FIRM) to serve as a pipe connecting the relevant companies and research institutions, both Japanese and foreign. FIRM’s Representative Director and Chairman Yuzo Toda stresses, “Japan, with its aging population, has a strong need for regenerative medicine, and it also has the technological strength to develop practical applications based on research results.”
According to FIRM, some foreign companies are now considering making Japan the initial market for their new regenerative medical products so as to achieve quick practical use for them. Toda declares, “Conditions are in place for realization of Japan-made, world- first products. FIRM’s role is to serve as an incubator for the regenerative medicine industry. We want to advance initiatives to open up this new field, working in tandem with Japanese and foreign companies.”
FIRM has established a partnership with the Alliance for Regenerative Medicine (ARM), America’s trade association for regenerative medicine. It also has tie-ups with Sweden, Britain, and Australia, and Canada will soon join the list. Japan’s initiatives in the field of regenerative medicine are developing international breadth.
Forum for Innovative Regenerative Medicine official website
Global scale of the regenerative medicine market
The future scale of the global market for regenerative medicine is forecast to grow to JPY 1 trillion (USD 9 billion) by 2020 and JPY 38 trillion (USD 345 billion) by 2050.
A medical field drawing global attention
Examples of regenerative medicine using iPS cells, embryonic stem cells, and somatic stem cells, now drawing attention for its potential in curing previously untreatable conditions.
Shortening of development period under expedited approval system
By comparison with the previous approval process, this system provides for a major shortening of the time required for approval of pharmaceuticals, subject to further confirmation after marketing, followed by a review of the approval.
Two regenerative medical products approved
Yuzo Toda, Representative Director, Chairman
Forum for Innovative Regenerative Medicine (FIRM)
FIRM, with 185 corporate members as of January 2016, is working to close gaps in awareness and regulations regarding regenerative medicine among industry, academia, government, and citizens, as well as to show specific ways forward.
Masayo Takahashi - "Hope - Yes" & "Patients First"
During ISSCR 2015 I had the opportunity to sit down with Masayo Takahashi of Riken. The interview can be found here and is an update to the below information on the first in human iPS clinical trial for Wet AMD.
Paul Knoepfler ran the interview and also provided a background story on the regulatory pause in Japan on the trial pending review of the data & application to move forward using a different cell source.
Masayo Takahashi: Leader of First Ever In-human Clinical Study Using iPS cells
Masayo Takahashi – Regenerative Medicine Using iPS Cells
RIKEN's iPS RPE cell sheet science for the Eye is being commercially developed by an arm's length technology Licensee, Healios. To-date the company has secured a number of important program partners and funding to establish a vertically integrated operation for global product marketing. Part of the strategy is to align the technology in international market regions - such as Europe and the US. Manufacturing and scientific collaborations are being established and structured to ensure strength (such as with UCL in the UK).
“Healios is seeking conditional approval for HLCR011 under newly established regulations for conditional and temporary approval (“conditional approval”) for cellular and tissue-based products (Article 23 and Article 26, Clause 1 of the revised [Japanese] Pharmaceutical Affairs Law)” https://www.healios.co.jp/en/development/pipeline/
"Healios entered into a joint development agreement with Sumitomo Dainippon Pharma Co., Ltd. in 2013. Under the agreement, Sumitomo Dainippon Pharma provides funds to Healios for co-developing iPS cell–derived human retinal pigment epithelial (RPE) cells and obtaining approval for the manufacture and sales of such products. Additionally, our two companies established a joint venture, SighRegen, in 2014 to promote the manufacture and sales of commercialized RPE cell products.
Healios is also working with Shibuya Kogyo CO., LTD., Nikon Corporation and Osaka University on the development of an automated culturing device with a view towards greater manufacturing efficiency. In such ways, we are building a strong development ecosystem and value chain so that we can promote the practical application of iPSC regenerative medicine products.
We are also planning for Healios-led development outside Japan. To date, we have completed the selection of manufacturing subcontractors in the US and European markets. We have also embarked on technology transfers to those subcontractors, and are in consultations with US regulatory authorities." https://www.healios.co.jp/en/development/
Riken will tap iPS cell bank for transplant NIKKEI Asian Review - March 21, 2015
OSAKA -- "A stockpile of stem cells derived from other people will be used in clinical trials of cell transplants for patients with an incurable eye disease, a potential breakthrough that could sharply reduce costs and waiting time.
Masayo Takahashi of the government-affiliated Riken research institute announced the plan Friday. Under a project led by her, a patient with age-related macular degeneration received a transplant of cell sheets made from induced pluripotent stem cells, or iPS cells, in September. The iPS cells were created from the patient's own cells to avoid transplant rejection.
For the second trial, slated to take place as early as 2017, iPS cells derived from other people will be used. While the first method is believed to have cost 50 million yen to 100 million yen ($410,600 to $821,300), Takahashi said that using stockpiled cells could reduce the cost to less than 10 million yen. And preparation time would be shortened from nearly one year to about six months.
"From a stock of iPS cells from one person, we can cultivate enough cells to transplant to dozens of patients," Takahashi said.
Kyoto University and others working to prepare a frozen stockpile of iPS cells for distribution will choose donors with special physical attributes unlikely to cause rejection.
Experiments on monkeys have yielded promising results. Little or no medication to curb rejection may be needed.
The iPS cell bank, with thorough control of safety and quality, is expected to help clear many hurdles to making regenerative medicine using stem cells accessible."
Foundation for Biomedical Research and Innovation RIKEN Press Release - September 12, 2014
First RPE cell sheet graft transplant
"Today, the first participant in the “Clinical study of autologous induced pluripotent stem cell-derived retinal pigment epithelium (RPE) cell sheets for exudative age-related macular degeneration (AMD)” underwent transplantation of a cell sheet graft. Surgical schedule Location: Institute for Biomedical Research and Innovation (Kobe, Japan) Start of procedure: 14:20, September 12, 2014 End of procedure: 16:20, September 12, 2014 Surgical staff: A team led by Yasuo Kurimoto, M.D. and two other ophthalmologists. Result A single RPE cell sheet (1.3 mm × 3 mm) was engrafted into the subretinal space of one eye. There was no serious hemorrhaging or complications. Research subject Residence: Hyogo Prefecture Age: 70s Gender: Female To protect patient confidentiality, and out of respect for the strong wishes of the individual, no information other than that given above will be made public. Research funding This clinical study is funded by the Japan Science and Technology Agency and the Ministry of Health, Labour and Welfare." http://www.riken-ibri.jp/AMD/img/20140912_1en.pdf
Project SummaryClinical Research:
Testing for safety and efficacy is required before any new medical intervention can enter routine use. Such clinical research takes various forms that differ in the number of human research subjects and features of the study design. In Japanese law, two main types of clinical experiments are recognized: registered clinical trials, which are used to test medical products prior to market authorization, and general clinical research, which includes smaller-scale studies, such as the pilot study described [...], and evaluations of some medical procedures, such as surgical techniques.
The research plan introduced [...] is for an open-label study of the transplantation of autologous induced pluripotent stem cell (iPSC)-derived retinal pigment epithelium (RPE) cell sheets in patients with exudative age-related macular degeneration (AMD). This is a very early-stage form of clinical research, and is intended to assess the safety of this intervention; it is not expected to yield significant improvements in visual acuity or other symptoms in the patients who participate in the study.
1) The current study is designed to evaluate this intervention only in patients with the exudative (wet-type) form of age-related macular degeneration.
2) The primary outcome to be assessed in this study is the safety of the intervention. If it is found to be acceptably safe, future studies will evaluate its efficacy. Patients who participate in this pilot study are not expected to experience dramatic improvements in their symptoms.
3) Enrollment in the study is limited to six subjects, as the safety of the intervention has not been established.
4) This pilot study is a very early stage of clinical research into the use of an iPSC-based intervention. Even in the event that this study suggests the intervention is acceptably safe to proceed to subsequent stages, a great deal of further research, including full-scale clinical trials, will be needed to more thoroughly evaluate the safety and efficacy of this approach.
Generation and Transplantation of RPE Cell Sheets:
In the first stage of the study, a 4 mm biopsy of tissue will be collected from the skin of each subject’s upper arm and cultured in a cell processing center. The skin cells will then be used to generate induced pluripotent stem cells (iPSCs). The autologous iPSCs will next be differentiated in culture into monolayer ‘sheets’ of retinal pigment epithelium (RPE) suitable for transplantation. The entire process, including regular assessments of safety and quality, will require approximately 10 months for each patient.
Once complete, the RPE cell sheets will be transplanted into the affected site in a single eye, from which the neovascular tissue has first been removed. Subjects will be monitored and followed up for four years following transplantation.
Potential Risks and Benefits:
The primary goal of this pilot study is to evaluate the safety of the transplantation of autologous induced pluripotent stem cell (iPSC)-derived retinal pigment epithelium (RPE) cell sheets in patients with exudative age-related macular degeneration (AMD). Patients who participate in this study are not expected to experience dramatic improvements in visual acuity or other symptoms. In the event that the intervention is found to be acceptably safe in this small group of patients, much additional research will be needed to evaluate its safety and efficacy in larger numbers of patients.
This is a first-in-human study using RPE cells differentiated from a pluripotent stem cell source. While intensive preclinical research in cultured cells and animal models has been conducted, it is impossible to rule out the risk of tumor formation triggered by the transplanted cells. The harvesting of tissue from the arm to be used for generation of iPSCs is an invasive procedure and will be conducted under anesthesia, both of which are associated with some risk, and the transplantation procedure is associated with risks including bleeding, infection, retinal detachment, any of which can lead to loss of visual function. The physicians conducting the study will monitor closely for any such adverse events, and take all appropriate measures should an adverse event occur.
After obtaining informed consent, volunteers will be screened for eligibility to participate in the study following a number of predefined inclusion and exclusion criteria – those selected will be enrolled in the ‘primary registration’ for the study. After primary enrollment, skin samples will be harvested from each patient and used to generate autologous iPSC-derived RPE cell sheets, a process that will take approximately 10 months.
After safety and quality testing, the RPE cell sheet will be prepared for transplantation. The terms of the original informed consent document will be reconfirmed with each subject, and providing the subject’s consent remains unchanged, he or she will be enrolled in the ‘secondary registration’ prior to transplantation. All patients will be intensively monitored for one year, with additional follow-up for three years following transplantation of the RPE cell sheet.
All subjects will be monitored intensively for the first year following the transplantation of the RPE cell sheet, with monthly evaluations for the first 6 months and bimonthly evaluations for the next 6 months. These evaluations will include tests of visual function, intraocular pressure, and medical imaging. Data from these examinations will be compiled and analyzed to evaluate the safety of the intervention and any effects on visual function. During the 3-year follow-up period, subjects will have annual eye examinations. Comprehensive cancer screening will be conducted at the harvesting of the skin biopsy, prior to the transplantation of the RPE cell sheet, at one year post-transplant, and at the end of the follow-up period (4 years post-transplant). Following this 4-year post-transplant monitoring and follow-up period, additional long-term observation of the study subjects will be periodically conducted.
This pilot study is a joint clinical research project coordinated and conducted by RIKEN, the IBRI (Institute for Biomedical Research and Innovation) Hospital, and the Kobe City Medical Center General Hospital. The IBRI Hospital will harvest skin biopsies, transplant RPE cell sheets, and conduct pre- and post-transplant monitoring of subjects. RIKEN will generate autologous iPSCs from the subjects’ skin cells and differentiate the iPSCs into RPE cell sheets. Kobe City Medical Center General Hospital will conduct some of the examinations during the monitoring period, support the screening process and, if necessary, provide emergency medical services.
The organizations involved in the current pilot study are urgently working to develop safe and effective treatments for age-related macular degeneration. This study is only the first step of many that will be needed to determine whether this transplantation of autologous iPSC-derived RPE cell sheets will be appropriate for use in the treatment of this disease. Due to the preliminary nature and small size of the study, the requirement for intensive long-term monitoring, the possible need for rapid access to emergency medical treatment, and the importance of clear communications with research and medical staff, including documentation required for the study, enrollment will be limited in principle to residents of Japan. The results of the study will be published as soon as they are available, and if they support the preliminary safety of this approach, may lead additional larger-scale research and development efforts in Japan and other countries.
About AMD - Retinal Pigment Epithelium
The macula is the central region of the retina and plays important roles in light-sensing and visual acuity. The retina is made of up a sensory retina layer that contains a type of neurons known as photoreceptors that convert light signals into neural activity and a supporting layer of non-neural cells called retinal pigment epithelium (RPE). The RPE acts as a barrier that separates the eye from the rest of the body, absorbs cellular waste materials, and also serves as a source of molecules called trophic factors that are needed for the survival and function of other nearby cells. Damage to the RPE can thus also affect the function of the sensory retina.
In age-related macular degeneration (AMD), the function of the macula deteriorates with age, which can be due to a number of causes. In the neovascular or exudative form of AMD (often called ‘wet-type AMD’), abnormal growth of new blood vessels occurs in the region immediately next to the eye (a process known as choroidal neovascularization). These new blood vessels may leak serum or blood and invade the overlying tissue, causing damage to the RPE and sensory retina.
The early symptoms of AMD include loss of acuity and contrast sensitivity in the center of the visual field, object distortion, blurring, and dimness of vision. In some cases, the onset of symptoms is rapid. As AMD increases in severity, it can lead to retinal detachment or hemorrhage, which can cause loss of vision in other parts of the eye as well.
The precise cause of AMD remains unknown, but aging, inflammation, and genetic factors are believed to play roles in weakening of the RPE. A second form of the disease, atrophic (or dry-type) AMD, is prevalent in people of European descent, while the wet-type is more common in Japan and other parts of East Asia. In Japan, around 1% of all people aged 50 and older suffer from some form of AMD.
Most current forms of treatment for wet-type AMD focus on stopping the abnormal blood vessel growth known as choroidal neovascularization, with the goal of preventing further progression of the disease. If neovascularization is absent in the fovea (the center of the macula), a laser can be used to ablate the blood vessels, while in cases of foveal neovascularization, agents that inhibit the activity of VEGF, a molecule that promotes the formation of blood vessels, have been approved for use. However, such treatments only prevent the progression of the disease, and cannot repair tissue damage that has already occurred to the RPE or sensory retina. Thus, therapies that both halt the growth of new blood vessels and reconstruct damaged tissue are needed to fully treat the effects of this disease.
RIKEN CDB 2-2-3 Minatojima-minamimachi, Chuo-ku, Kobe 650-0047, Japan
Foundation for Biomedical Research and Innovation
2-2 Minatojima-minamimachi ,Chuo-ku, Kobe 650-0047, Japan
Copyright 2013 RIKEN-Foundation for Biomedical Research and Innovation. All rights reserved.
- Transplantation of Embryonic and Induced Pluripotent StemCell-Derived 3D Retinal Sheets into Retinal Degenerative Mice – Stem CellReports, May 2014
- Characterizationof Human Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium CellSheets Aiming for Clinical Application – Stem Cell Reports, Feb 2014
- Tumorigenicity Studies of Induced Pluripotent Stem Cell(iPSC)-Derived Retinal Pigment Epithelium (RPE) for the Treatment ofAge-Related Macular Degeneration – PLOS One, Jan 2014
- Protective effects of human iPS-derived retinal pigmented epithelial cells in comparison with human mesenchymal stromal cells and human neural stem cells on the degenerating retina in rd1 mice – Stem Cells, Feb 2015
- In vitro and in vivo characterization of pigment epithelialcells differentiated from primate embryonic stem cells. PubMed, Mar 2004
RIKEN Eye Patent Portfolio (representative Eye IP only):
- METHODS FOR PRODUCING RETINAL TISSUE AND RETINA-RELATED CELL/METHOD FOR PRODUCING RETINAL PIGMENT EPITHELIAL CELL / METHOD FOR PRODUCING RETINAL TISSUE / METHOD FOR PRODUCING OPTIC-CUP-LIKE STRUCTURE / METHOD FOR PRODUCING RETINAL-LAYER-SPECIFIC NERVE CELL