On January 20th 2014 the Cure Girls went to visit the labs of pharmacology of the university of Milan and met with Dr. Daniele Bottai. He showed us the new labs and gave us an interview to explain what his team is working on with regards to a cure for SCI.
1. Can you briefly describe the research that you are doing in relation to spinal cord injury?
When in 2006, I moved at the University of Milan, I have begun to get interested in Spinal Cord Injury (SCI) using a new (at the time) “drug”: neural stem cells. Between 2002 and 2006, I worked in the Laboratory of Professor Angelo Vescovi where I learned to manipulate neural stem cells (from different regions of the brain) both human and mouse.
In these 8 years spent to the University of Milan we have been studying the role of different types of stem cell in transplantation in animal model of SCI, in particular, we have studied the effects of murine embryonic and neural stem cells and human amniotic fluid (AFCS ) with the purpose to find the sources of stem cells that were an available source and with the appropriate characteristics for the treatment of neurological diseases .
In general, we can say that these cellular processes (performed in acute spinal cord lesion) have positive effects and are significant from a functional and morphological point of view. After treatment with the cells listed above, the mice returned to walk (albeit not the same as they did before the lesion); while lesioned not treated animals are able to move their hind limbs, but not to walk. (These results have been summarized in three scientific papers:
D. Bottai, D. Cigognini, L. Madaschi, R. Adami, E. Nicora, M. Menarini, A.M. Di Giulio, A. Gorio (2010).Embryonic Stem Cells Promote Motor Recovery and Affect Inflammatory Cell Infiltration in the Spinal Cord Injured Mice Experimental Neurology 223, 452-463 ;
D. Bottai , L. Madaschi, A.M. Di Giulio and A. Gorio. (2008) Viability -Dependent Promoting Action of Adult Neural Precursors in Spinal Cord Injury . Molecular Medicine , 14 (9-10), 634-644.
On the bases of these results, we asked what was the mechanism that caused this improvement.
The answer was that in this model the role of animal cells is purely trophic and they are not going to replace, if not in small portion, damaged or dead cells.
Various are the trophic molecules (cytokines) that are involved in this phenomenon. Recently, we have focused our attention on amniotic fluid cells. We chose this cell type because their availability since the at term cesarean delivery could represent an unlimited source of stem cells with no ethical issues and few risks for the child and the mother.
In a work that a few days ago has been accepted for publication (D. Bottai , G. Scesa , D. Cigognini, R. Adami , E.. Nicora, S. Abrignani, A.M. Di Giulio, and A. Gorio Third trimester amniotic fluid NG2 -positive cells are effective in improving on repair in spinal cord injury. Experimental Neurology.) we have shown that a trophic factor, produced by the AFCS, which is important for the induction of the morphofunctional recovery, was the hepatocyte growth factor (HGF) and that this cytokine was produced only by particular sub- populations of our cells or those expressing on their surface the NG2 protein (a membrane proteoglycan). Such a membrane protein can be hopefully used in the future to select from the amniotic fluid liquid the cells that express NG2 and so have a therapeutic action.
We are currently investigating what is the correlation between NG2 and HGF.
2. Acute injury or chronic injury present any difference for a research approach? Could you explains the differences and the advantages and disadvantages.
I do not think we can talk about differences between acute and chronic lesion in terms of advantages and disadvantages. These are two different pathological conditions, the acute progress into the chronic with the passage of time mostly because there is a de- myelination process.
In this context, we are dealing with two different types of patients the acute ones, namely that a few days or weeks have suffered damage to the spinal cord which have a very extensive inflammatory condition that exacerbates the primary mechanical injury further damaging the tissue and those, who instead, underwent chronic spinal cord injury for more time (months and years) in which degeneration induced by primary damage and the secondary SCI causes the formation of a cavity surrounded by the glial scar that separates the lesion from the undamaged tissue and prevent nerve regeneration.
Currently the researcher and clinician are faced with these types of patients because in the first instance they have not been able to prepare effective therapies to treat acute patients.
The therapeutic approach to the patient who recently underwent spinal damage is intended to reduce the compression state and to control the secondary damage due to inflammation through the drug methylprednisolone, inter alia, that approach does not seem to have a sufficient efficacy, as evidenced by the fact that the number of chronically para or quadriplegics is unfortunately increasing.
In the chronic patient instead we find ourselves facing a very different situation with the blood-brain barrier that is closed, and a glial scar consisting mainly of fibroblasts from the meninges and reactive astrocytes that produce proteoglycans (extracellular matrix molecules) that are responsible for the inhibition growth of axons.
In this situation, the therapeutic approach is vastly different from that prepared in the state of acute spinal cord injury.
In fact, removal (either mechanical – surgical or enzymatic) is a sine qua non con-diction in order to prepare any kind of intervention to restore or replace dead or damaged cells and rebuild the axons making them grow in the appropriate direction.
In this context, the treatment of chronic patients need multiple concurrent interventions:
- Treatment with drugs that induce axonal regeneration;
- Treatment with drugs that reduce the inhibitory effects of glial scar both mechanical and enzymatic chondroitinase that due to factors such as chemical inhibitors, blockers of Nogo and other myelin components;
- Treatment with cells or systems consisting of nanomaterials and cells.
Some of these approaches were ineffective few years ago but in the light of developments in nanomaterials and new types of stem cells I think it might be appropriate to re-examine these pathways.
3. How do you think we can solve the problem of scar?
As I mentioned in the previous answer, in order to find an approach that leads to the recovery of sensory and motor pathways, it is necessary to make the scar area permissive for the survival of cells that are transplanted and that would allow axonal growth in the manner to ensure the recovery of the routes of transmission.
With this in mind, scar removal is definitely needed and should be prepared to minimize the risk of inducing further damage.
In this context, the experimental work in the preclinical phase or with animal models is essential but at the same time very complex given that small animals have practical difficulties of intervention and larger animals have problems is housing costs that are beyond the economical capability of most the laboratories that I know.
Finally, the translation of the results obtained in the preclinical stage is very difficult for a variety of patients such as those with spinal cord injury whose disease is highly variable due to the fact that the damage is very random and therefore leads to differences between the patient and the other.
4. Can you apply your research to Chronic Spinal Cord Injury?
The applicability of the cells in the amniotic fluid in models of chronic spinal cord injury must be verified experimentally, so I can say a priori that such an intervention can be prepared but obviously need the appropriate adjustments of the experimental protocol. In fact, while in the acute model we have a purely trophic action in the case of chronic treatment the intervention should be at the level of local scar in order to determine whether these cells could contribute to modify the scar itself and reconstitute the ways by means of the differentiation in cells central nervous system (neurons, oligodendrocytes and astrocytes) or by inducing endogenous stem cells to differentiate into mature cells. In this context, previous treatment with chondroitinase could improve the success of the experiment.
5. Do you have collaborations with other research institutions ? Which ones?
As I mentioned in our discussion in the institute , I believe that partnerships are the lifeblood of research. In recent years I have had collaborations with various research groups and consortia. Firstly put the FUNGENES: Functional Genomics of Human Embryonic Stem Cells; funded by the European Economic Community, Sixth Framework Programme. (€ 500,000 for 3 years) (in collaboration with Prof. A.L. Vescovi of which I was the deputy). This project was set out to investigate the characteristics proliferative and differentiative of stem cells (especially embryonic). This project involved and brought together about a dozen institutions across Europe and basically was the first step that allowed me to improve my knowledge on stem cells.
– Study of functional recovery induced by transplantation of neural stem cells in animal models of acute spinal cord contusion, funded by Fondazione Cariplo.(€ 300,000 for 2 years) (Coordinator Prof. A.L. Vescovi).
It was a project that introduced me in the world of spinal cord injury, and thanks to Prof. Vescovi, the project that make me chose to continue the research in neurodegenerative diseases
– Neural stem cells: a new approach to mobile spinal muscular atrophy; Asamsi non-profit organization funded by foundations and Families of SMA Italy . (Head with Prof. A. L. Vescovi) .
I am also currently working on the project “Role of stem cells in the treatment of glaucoma” (glaucoma is a neurodegenerative disease) with Professor Mario Luca Rossetti, which conducts clinical and research in my department and with Dr. Valentina Massa (which also works in my department) for a study of neurological disorders.
6. What are the steps needed to translate your result in human?
To start a clinical trial phase 1-2, that provide the safety analysis (which most likely this type of cells have since they belong to the class of mesenchymal which have already been extensively tested in several clinical trials) and effectiveness, our results must be first validated in other laboratories. Subsequently, it will be necessary to derive the cells so that they are compatible with the transplant in humans that means that the cells must satisfy conditions of Good Manufacturing Practice (GMP ) that involves the use of materials “human grade” in order to reduce the risk to the patient (for pathologies as the well- known prion disease such as mad cow disease. This procedure is currently out of our economical availability as it requires economical conditions of sterility and purity that we cannot get unless you build the appropriate laboratories.
7. Is there any particular obstacle that slows down your work?
The current financial situation of our country, where the cuts have affected many strategic sectors of the economy and cultural is experienced by us researchers, with much apprehension. While it is true that funding should be allocated to those who do the research and then excellence must be a prerequisite for this contingency (Italian and international), cuts in recent time unfortunately affected groups or researchers that produce high quality work. So there is now the hope that this trend may change and that organizations and associations (onlus) can help the researcher by funding specific projects.
We thank Dr. Bottai for his precious work and for giving us the opportunity to visit the center and for answering our questions.
Cure Girls Arcangela, Marina and Loredana
I desagree with what is said in the answer #2 as I believe that by comparing acute spinal cord injury (SCI) research with chronic it is possible to show several advantages and desadvatages at different levels (scientific, economic, etc) that are rarely considered.
I would like to point out first that if we had a cure for chronic SCI all people living with SCI could benefit from it as even someone that will get SCI tomorrow will become chronic in a few weeks. The opposite is not true as usually a therapy that works for acute SCI can be effective just if applied within few hours or at best in a few days after injury. For this reason it would be logical to give priority to SCI research that focus on chronic SCI.
Unfortunately so far SCI research has been mostly acute oriented and, in my opinion, that shows a luck of interest from researchers toward people living with SCI, but when they look for funding.
In fact often people with SCI raise money and donate to SCI researchers that then in most cases will do acute SCI research without making any ethical consideration.
It’s hard to believe, but that is what I have seen happening many times.
From a more practical perspective to study rodents with chronic SCI takes at least 6 weeks longer as animals after SCI need at least 6 weeks to become chronics and then get the experimental therapy. So researchers need to work 6 weeks longer with the associated costs to conclude a study and hopefully get it published in a scientific journal. That will help them get another grant to do another study and so on. Clearly for a researcher it is easier and more convenient to do just acute studies.
In case a study produces really good results and it is decided to test the therapy on people a lot of money are necessary to complete clinical trials and in most cases it is necessary to find investors that will take the risks and the costs to conduct clinical trials to then eventually make profit if the therapy is proven effective and made available to all the patients. Very often it happens that the costs of clinical trials are too high in comparison to the potential profit opportunity therefore it becomes impossible to find investors and the therapy ends up in the so called “valley of death”.
Now let’s consider the case we have a therapy that might be effective in people with chronic SCI and needs to be taken to clinical trials.
First the number of patients that could benefit from it is much higher, so is the potential profit. That makes it easier to find investors.
Then a clinical trial on people with chronic SCI has several advantages, for example:
1) Patients can be recruited for the existing population living with chronic SCI. That makes it faster to recruit the patients needed which makes the trial faster and cheaper than an acute trial.
2) People with chronic SCI can travel more easily so the trial can be conducted in just one center further reducing costs
3) A patient with chronic SCI is neurologically stable, therefore can be used as his own control to determine the effects of the therapy (no need of a control group). So fewer patients are needed to complete the trial which result in further saving of time and money.
Conclusion, acute SCI research very often has no chance to be translated into humans. So why is most SCI research acute focused? It seems very possible that the reason is that researchers reputation and their career depends on the number of publications they produce, not on how benefical their work is for people and the society. So why should they bother working on chronic SCI since it’s a bit more complicated for them?
I believe that the way SCI research is being funded should be totally reconsidered and the focus should be moved on chronic SCI.