1 Stem Cell Therapy Applications in pediatric neurology Ali Fatemi, MD Director of Neurogenetics Division Kennedy Krieger Institute Johns Hopkins University Baltimore, USA The Author has nothing to disclose in relevance to this lecture
2 HYPE OR HOPE??
3 History of stem cell transplantation
4 LEARNING OBJECTIVES/OUTLINE THE TARGET DISEASE DEFINE STEM CELLS AND DIFFERENT CELL TYPES REVIEW MECHANISMS OF ACTION IN CELL-BASED THERAPY REVIEW CURRENT CLINICAL TRIALS REVIEW POTENTIAL ADVERSE EVENTS
6 Rule of thumb for therapies Each condition will require targeted treatment. Stem cells will not cure all diseases. If a disease is static, regenerative therapy is much more likely to be effective than if a disease is progressive. Need for surrogate markers and proper controls to evaluate therapies. DO NO HARM! Need for careful monitoring for adverse events which may outweigh benefits.
7 TIMING AND MODE OF INTERVENTION IS DISEASE SPECIFIC Age of patient (changes in immune response) Time of disease onset (more likely to be effective if treatment is early to prevent destruction) In rodent studies for different disease models different time points have been determined, but extrapolation to humans very difficult
8 LEARNING OBJECTIVES/OUTLINE THE TARGET DISEASE DEFINE STEM CELLS AND DIFFERENT CELL TYPES REVIEW MECHANISMS OF ACTION IN CELL-BASED THERAPY REVIEW CURRENT CLINICAL TRIALS REVIEW POTENTIAL ADVERSE EVENTS
9 Defining stem cells Stem cell: unspecialized cell that is capable of replicating itself but also differentiate into specialized cells.
10 Different cell types Human body after birth has > 200 cell types All cells derived from embryonic stem cells Organs also contain stem cells in niches referred to as somatic stem cells.
11 Embryonic stem cell
12 Myeloid derived stem cells Hematopoietic stem cells -CD34+ cells can give rise to all blood cell types, few controversial reports that under certain conditions in-vitro these cells can become neurons Mesenchymal stem cells can be differentiated to a variety of different tissue cells in-vitro including neurons. 50,000 adult and pediatric patients/year world-wide have received bone marrow stem cells.
14 Neural Stem Cells Neural stem cells can be derived from adult and fetal human cadavers Mouse embryonic stem cells can be differentiated to neural stem cells in-vitro Neural stem cells have the ability to become neurons, astrocytes, oligodendrocytes and endothelial cells. Neurons Embryonic Stem Cells NSC NRP APC Astrocytes Somatic Stem Cells (Fetal or Adult Tissue) GRP OPC Oligodendrocytes
15 Glial Precursor Cells
16 Traditional Sources of Stem cells
17 Induced Pluripotent Stem Cells (ipsc) Shinya Yamanaka Nobel Prize 2012 in Physiology or Medicince
18 Induced Pluripotent Stem Cells (ipsc) cells that have been genetically reprogrammed to an embryonic stem cell like state initial method based on using 4 different lentiviral vectors containing OCt4, Sox2,Klf4, c-myc. Mouse ipscs were first reported in 2006, and human in late ipscs demonstrate important characteristics of pluripotent stem cells, including expressing stem cell markers, forming tumors containing cells from all three germ layers
19 ips cells make viable mice Tetraploid Complementation Assay From Zhao XY et al. Nature Protocols 5, (2010)
20 OUTLINE THE TARGET DISEASE DEFINING STEM CELLS AND DIFFERENT CELL TYPES MECHANISMS OF CELL-BASED THERAPY Clinical trials Potential Adverse Events
21 How exogenous cells may help 1. Restore tissue by becoming neurons or glial cells and integrate into the neuronal network 2. Restore tissue by promoting activation of endogenous stem cells 3. Alter inflammatory response Unlikely to have an effect in brain malformations (schizencephaly, lissencephaly, Holoprosencephaly etc.)
22 Restoration by Differentiation Embryonic, neural and CD133+ stem cells are all thought to be able to differentiate into neurons when transplanted directly into the CNS in rodents
23 Nature Reviews Neuroscience 2002, 3; 401
24 Neural Spheres migrate and stimulate endogenous stem cells
25 INTEGRATION EXAMPLE Human Glial Precursors remyeliante the shiverer Windrem MS et al. Nat Med 2004
26 Glial precursors migrate and differentiate along the white matter
27 Limited co-localization of GRPs with astrocytes at 1 month GFAP GFP Fatemi A. et al. 2012
28 Glial precursors migrate and differentiate along the white matter
29 GRPs differentiate into mature Oligodendrocytes at 2 months Fatemi A. et al. 2012
30 How exogenous cells may help 1. Restore tissue by becoming neurons or glial cells and integrate into the neuronal network 2. Restore tissue by promoting activation of endogenous stem cells 3. Alter inflammatory response Unlikely to have an effect on defects of brain formation or neuronal migration
31 Trophic effects Many studies show that transplanted animals often have better outcome even if no transplanted cells are found in the brain. Many stem cells release growth factors have immuno-modulatory effects.
32 Mesenchymal stem cell injection improves outcome in mice with neonatal hypoxia-ischemia Cells injected into the brain 3 days after HI Van Velthoven et al. Brain, Behavior, and Immunity Volume 24, Issue 3, March 2010, Pages
33 Mesenchymal stem cell injection increases endogenous stem cell proliferation after neonatal hypoxia-ischemia Cells injected into the brain 3 days after HI Van Velthoven et al. Brain, Behavior, and Immunity Volume 24, Issue 3, March 2010, Pages
34 How exogenous cells may help 1. Restore tissue by becoming neurons or glial cells and integrate into the neuronal network 2. Restore tissue by promoting activation of endogenous stem cells 3. Alter inflammatory response Unlikely to have an effect on defects of brain formation or neuronal migration
36 OUTLINE THE TARGET DISEASE DEFINING STEM CELLS AND DIFFERENT CELL TYPES MECHANISMS OF CELL-BASED THERAPY CLINICAL TRIALS Potential Adverse Events
37 Bone marrow stem cell transplantation Has improved outcome in some neurometabolic disease High risk of mortality (5-30%) pending on patient s age, baseline disease and other factors High morbidity due to chemotherapy and immunosuppressive treatment leading to infections and organ damage.
39 Br J Haematol Feb;148(3): Epub 2009 Nov 16.
40 Cord Blood nucleated cell transplantation in neurologic diseases Recently used in neurometabolic disease. Associated with lower morbidity and mortality, easier to find a matched donor Role of autologous transplantation
41 Autologous Cord Blood Transplantation in Cerebral Palsy More than 170 children with diagnosis of cerebral palsy have received autologous CB units as compassionate care at Duke University Ongoing Trial in USA, NCT , Inclusion Criteria: 1-12 year of age, placebo-controlled, FU after 1 year Clinical evidence of a non-progressive motor disability due to brain dysfunction. The subjects will not have the ability to sit independently by one year of age or the ability to walk by 18 months of age. 2 other studies: Korea (allogeneic + Epo) and Iran (CD133+ autolog)
42 Autologous Cord Blood Transplantation in HIE 2 Ongoing trials listed at clinicaltrials.gov: NCT (Duke Univ. USA) Term Infants with moderate to severe encephalopathy that missed cooling receive autologous cord blood within first 14 days Compared to historical controls Outcome will be assessed at 9-12 months NCT (Mexico) Term Infants with HIE within the first 48h of life Compared to controls who refuse therapy Outcome at 1 year
43 Phase I (safety) trial of Neural Stem Cells in USA First FDA approved study of human fetal tissue derived neural stem cells, was conducted by a biotech company In 6 patients with advanced stages of infantile and late infantile Neuronal Ceroid Lipofuscinosis due to PPT1 deficiency. Study completed in 2008, data presented as poster no peer reviewed publication yet Cells directly transplanted into different sites into the brain via stereotaxic surgery, 3 patients got 500mil cells, 3 got 1bil cells Patients immunosuppressed for 12 months Monitored for 4 years, 2 patients died presumably due to disease progression (autopsy: no sign of toxicity), others tolerated treatment well but all progressed. Autopsy showed transplanted cells, no details published.
44 Phase I Trial of Neural Stem Cell in a Leukodsytrophy in USA conducted at a major academic institution in US in collaboration with a Biotech company enrolled 4 patients with genetically confirmed Pelizaeus- Merzbacher Disease immunosuppression for 9 months Cells again directly transplanted into different sites into the brain via stereotaxic surgery To be monitored first for 12 months intensely and then for 4 years. All patients have been enrolled, report of myelin signal in MRI around the injection site
45 OUTLINE THE TARGET DISEASE DEFINING STEM CELLS AND DIFFERENT CELL TYPES MECHANISMS OF CELL-BASED THERAPY Clinical trials POTENTIAL ADVERSE EVENTS
46 Considerations for Cell Based Therapy Stem Cells capable of forming tumors! Higher likelyhood with more immature cells like embryonic stem cells and neural stem cells Between 5-30% incidence in animals Comi A.M., et al., Pediatric Neurology 2008
48 Complications of hematopoeitic cell transplantation Acute and chronic graft versus host disease Engraftment syndrome Veno-occlusive disease Idiopathic pulmonary syndrome Immunosuppression associated infection Side effect of chemotherapeutics (preparative regimen)
49 Graft versus Host disease Graft cells attack body, very common in bone marrow transplant Acute: Within 3 months Diarrhea Liver disease Rash Fever in 19-66% of matched recipients Chronic: After 3 months Hairloss Rash Lung and digestive disease Liver disease Approximately one third of patients with matched recipient
50 Cell Based Therapy for Parkinson Disease LESSONS LEARNED In 1987, a series of small open-label trials, in which only carefully selected patients were included, was initiated human fetal dopaminergic neurons survived and function for more than 10 years in the striatum of patients with PD with benefits in some patients. 2 double-blind sham-surgery controlled trials were launched in the early 1990s turned out to be more disappointing than expected: showing only modest efficacy was observed in the transplantation groups, a significant number of patients developed worsening dyskinesias after transplantation.
51 Lessons learned from Krabbe disease the big thing in 2005
52 Longterm Follow Up study of same cohort published in presymptomatic children transplanted at Duke and elsewhere for early infantile Krabbe disease. 2 died All others spastic 3 mild 5 required gastostomies All were below 3% with height and weight All have abnormal expressive language 50% walk with assistive devices 25% don t walk Duffner P et al. Genetics in Medicine (2009) 11, 450
53 Non-academic institutions worldwide currently offering stem cell therapy Often many different disease are grouped into terms such as cerebral palsy No immunosuppressive therapy is discussed Almost all claim that there are no side effects Cell source and quality not clear No peer-reviewed publications of their claims High costs for patients
54 Summary Currently ongoing cell based therapy for a number of neurometabolic conditions Advanced cell engineering methods open the door to new therapeutic approaches Expect to have many more trials to come within the next 5-10 years Not all patients with Cerebral Palsy will be ideal candidates. Stem cell therapy can be harmful, very dangerous complications including death. Need for careful investigations to determine who will benefit and in whom it may be harmful.
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