Advanced Therapies in Neuroprotection and Immune-regulation

Home departments Celular Therapy and Regenerative Medicine Advanced Therapies in Neuroprotection

Research areas:

     Basic disease-oriented research: Characterization of the mechanisms of immune regulation, neuroprotection, and neurodegeneration of bioactive peptides in inflammatory/autoimmune disorders. Therapeutic exploitation of peptide-functionalised, nano-based drug-delivery systems in inflammatory/ autoimmune disorders. Activation of endogenous regulatory immune cells as an approach towards cell therapy. Immunologic effects and mechanisms of disease-linked protein aggregation and chaperone function.

     Patient-oriented research: Studies in patients suffering multiple sclerosis aimed to produce research results in profiling studies (such as metabolomics, gene or antigen arrays) to understand disease mechanisms from a systems biology perspective as a non-invasive approach to diagnose and grade neurodegenerative disorders with an autoimmune/inflammatory component. This could allow the assessment of new therapies during clinical trials, the identification of patients at risk to develop adverse effects during treatment, and the final implementation of new tools towards a more personalised medicine.

Basic immunology and translational research

Our immune system constantly interacts with our internal environment, protects us from our external environment and provides the inherent knowledge to sense the difference between friend and foe. Therefore, the immune system is one of the most dynamic body components in determining our state of health or disease. The importance of immune-based disorders is highlighted by the fact that immune-mediated inflammatory diseases are known to affect nearly 20 percent of the U.S. population. These conditions, which include juvenile-onset diabetes, rheumatoid arthritis, multiple sclerosis, asthma, systemic lupus erythematosus, and acquired immune deficiency syndrome, lead to chronic debilitation, life-long illness, protracted morbidity and result in great medical costs. The immune system also has a major involvement in post-transplant rejection and in the body’s ability to fight invading cancer cells.

Despite of these seemingly unconnected diseases, basic immunologists are in a prominent position to apprehend a broader picture under a cohesive paradigm. This forefront of future therapeutic strategies relies on the successful applications of genetic, molecular, and cellular immunology to prevent, treat, and cure the problems caused by immune-based disorders. The translational aspects of research that are directly related to patient care require integrated, multidisciplinary strategies involving teams skilled in basic biology and clinical investigation, ethics and regulatory issues. To take full advantage of such a global approach, it is essential to focus research programs on a limited number of well-validated basic concepts common to autoimmunity, allergy and organ transplantation. Thus, how to best enforce or end immune responses for the benefit of the patients is probably the most exciting attainable goal for today’s therapeutics.


Lines of research

    -Modulation of innate and adaptive immunity by Activity-dependent Neuroprotective
     Protein (ADNP) and ADNP-derived peptides.
    -Chemical synthesis and biological characterization of metal-protected nanoparticles
     functionalized with bioactive peptides.
    -Immunologic effects and mechanisms of disease-linked protein aggregation.


Modulation of innate and adaptive immunity by Activity-dependent Neuroprotective Protein (ADNP) and ADNP-derived peptides.

The discovery and characterization of new molecules and processes that regulate key activities on immunocompetent cells is of interest, but this is it especially when the same molecule has both neuroprotective and immunomodulatory activities. We found that Activity-dependent Neuroprotective Protein (ADNP) has a role in the immune homeostasis of the Central Nervous System (CNS). Moreover, ADNP is known to harbor neuroprotective activities that map to the ADNP-derived sequence octapeptide called NAP (NAPVSIPQ). Cellular and molecular mechanism of action of NAP and ADNP are being investigated including its potential in animal models of rheumatoid arthritis, experimental autoimmune encephalomyelitis, acute neuroinflammation, and inflammatory bowel disease. NAP remarkable pharmacological characteristics in terms of drug-discovery programs: a) intranasal, subcutaneous, intravenous, and intraperitoneal routes of administration, b) no side effects in toxicological studies, and c) cellular protective effects not linked to mitogenic activity. ADNP was initially characterized as a gene inducible in astrocytes by treatment with VIP, our current work demonstrates that ADNP is functionally expressed in the immune system as a VIP-responsive gene. The suppressive activities of NAP on innate and adaptive immunity, suggest that ADNP might mediate some of the immunomodulatory effects attributed to the cytokine-neuropeptide vasoactive intestinal peptide (VIP).


Chemical synthesis and biological characterization of metal-protected nanoparticles functionalized with bioactive peptides.

Engineering nanoconjugates is an extensive field of research due to its potential translation for biomedical applications such as probes for electron microscopy to visualize cellular components, drug delivery, detection, diagnosis and therapy (targeted and non-targeted). Among the different types of nanoparticles, noble metal ones are of special relevance due to their plasmonic properties -which make them well suited as molecular markers in sensitive light transmission and scattering measurements- together with their enhancement effect in Raman (SERS) or surface-enhanced infrared (SEIR) spectroscopy. There is also a growing interest in their biological features as nanocarriers of chemotherapeutic agents or by themselves, i.e., as potential cytostatic agents in cancer treatment, anti-angiogenic properties, immune regulatory effects or due to toxicity and/or environmental concerns. An important limitation to the therapeutic use of endogenic peptides is their short half-life due to the attack of endopeptidases. An efficient way of protecting peptides of biomedical interest from endopeptidases consists in their covalent binding to nanoparticles, also as it is the case for vasoactive intestinal peptide (VIP), we can take the advantage of a smart delivery of drug compounds to cancer cells that overexpress VIP receptors. Also, based on the immune properties of VIP, we expect to combine indirect and direct approaches, based on immunotherapy and chemotherapy.

The lab is part of NanoSpain, the Spanish nanotechnology network.





Immunologic effects and mechanisms of disease-linked protein aggregation.

Protein conformational diseases include a range of degenerative disorders in which specific peptides or proteins misfold and self-assemble into amyloid fibrils –characterised by a cross beta-sheet structure- that may then be deposited in specific tissues, resulting in cell death. These disorders include Alzheimer’s, Parkinson’s (PD), and Huntington’s diseases, and type II diabetes. Recently, work in mammalian cells and in model organisms has established a firm link between chaperone function and the panoply of conformational diseases and amyloidosis. However, the molecular mechanisms underlying such anti-aggregation properties have not yet been elucidated. Our research interest is to understand the biophysical basis for protein aggregation of alpha-synuclein and its structure-linked toxicity and activation of the immune responses. We plan to generate novel and more efficient functions in chaperone proteins by in vitro evolution as a way to improve their inhibitory aggregation features for biotechnological applications.


Patient-oriented research

Our interest is focused in multiple sclerosis (MS) patients and animal models of neuroinflammation. Besides the information of the immune response itself, we are interested in pattern recognition analysis. The goal is to provide two distinct but interlinked informational sets. On the one hand, the ability to differentiate between groups of samples is relevant by itself, especially in the clinical or pre-clinical environment. On the other hand, they also provide with functional information, and this information can be related to the physiopathology of MS. We use Magnetic Resonance Spectroscopy (MRS) evaluation techniques of metabolic parameters and gene expression profiling to perform in vivo studies in mouse models and human patients. It is possible to measure several different molecules and parameters by using either endogenous (for example, brain function could be related to the redox state of iron in deoxyhemoglobin) or exogenous contrast agents (31P-, 13C-, 1H-MRS). Always working in network, we have now the opportunity to implement those techniques both in the experimental settings to understand the pathology of the disease and its evolution and in the clinical (MS diagnosis and treatment follow-up) environments.

Those approaches relied on a fruitful collaboration with the team led by the clinical neurologist Dr. Guillermo Izquierdo and his team at the Virgen Macarena University Hospital, Sevilla.


Funding

   Federation of European Biochemical Societies (FEBS)
   Spanish Ministry of Health (Instituto de Salud Carlos III)
   Spanish Ministry of Science and Innovation
   Spanish National Cooperative Research Network in Multiple Sclerosis (RETIC-REEM)
   Junta de Andalucía (Consejeria de Innovacion, Ciencia y Empresa)
   Junta de Andalucía (Consejeria de Salud)
   University of Seville
   Fundación Salud 2000 Merck-Serono
   Fundación Alicia Koplowitz


Selected Publications:

C. Roodveldt, A. Labrador-Garrido, E. Gonzalez-Rey, R. Fernandez-Montesinos, M. Caro, C.C. Lachaud, C.A. Waudby, M. Delgado, C.M. Dobson, D. Pozo. Glial innate immunity generated by non-aggregated alpha-synuclein in mouse: differences between wild-type and Parkinson's disease-linked Mutants. PlosOne (In press). 2010.

R. Klippstein, D. Pozo. Nanotechnology-based manipulation of dendritic cells for enhanced immunotherapy strategies. Nanomedicine: Nanotechnology, Biology, and Medicine (Epub ahead of print). DOI 10.1016/j.nano.2010.01.001. 2010 

R. Klippstein, R. Fernandez-Montesinos, PM. Castillo, AP. Zaderenko, D. Pozo Silver nanoparticles interactions with the immune system: implications for health and disease In “Silver Nanoparticles" ISBN 978-953-028-05. Chapter 16. 309-324. 2010  IN-TECH Books. Vienna, Austria

R. Fernandez-Montesinos, M Torres, D. Baglietto-Vargas, A. Gutierrez, I. Gozes, J Vitorica, D. Pozo. Activity-dependent neuroprotective protein (ADNP) expression in the amyloid precursor protein/presenilin 1 mouse model of Alzheimer's disease. Journal of Molecular Neuroscience 41, 114-120. 2010

D. Pozo, P. Anderson, E. Gonzalez-Rey. Induction of alloantigen-specific human T regulatory cells by vasoactive intestinal peptide. Journal of Immunology 183, 4346-59. 2009

C. Roodveldt, CW. Bertoncini, A. Andersson, A.T. van der Goot, S-T D. Hsu, R. Fernandez-Montesinos, J. de Jong, T.J. van Ham, EA.A. Nollen, D. Pozo, J. Christodoulou, CM. Dobson. Chaperone proteostasis in Parkinson’s disease: stabilization of the Hsp70/?-synuclein complex by Hip. EMBO Journal. 28, 3758- 3770. 2009

R. Fernandez-Montesinos, PM. Castillo, R. Klippstein, E. Gonzalez-Rey, JA. Mejias, AP.  Zaderenko, D. Pozo. Chemical synthesis and characterization of silver-protected vasoactive intestinal peptide nanoparticles. Nanomedicine. 4, 919-930. 2009

JL. Herrera, E. Gonzalez-Rey, FJ Quintana, R. Fernandez-Montesinos, R. Najmanovich, D. Pozo. Toll-like receptor stimulation differentially regulates vasoactive intestinal peptide type 2 receptor in macrophages. Journal of Cellular and Molecular Medicine 13, 1-9. 2009

P M. Castillo, JL Herrera, R. Fernandez-Montesinos, C. Caro, AP.  Zaderenko,  J.A. Mejías, D. Pozo. Tiopronin monolayer-protected silver nanoparticles modulates interleukin-6 secretion mediated by Toll-like receptor ligands. Nanomedicine 3, 627-635. 2008

Pozo, D. Immune-based disorders: the challenges for translational immunology. Journal of Cellular and Molecular Medicine 12, 1085-1086. 2008

C. Roodveldt, J. Christodoulou, CM Dobson Immunological features of alpha-synuclein in Parkinson's Disease. Journal of Cellular and Molecular Medicine 12, 1820-1829. 2008

Pozo D, Vales-Gomez M, Mavaddat N, Williamson SC, Chisholm SE, Reyburn H. CD161 (human NKR-P1A) signaling in NK cells involves the activation of acid sphingomyelinase. Journal of Immunology 176, 2397-2406. 2006

Quintana FJ, Zaltzman R, Fernandez-Montesinos R, Herrera JL, Gozes I, Cohen IR, Pozo D. NAP, a peptide derived from the activity-dependent neuroprotective protein, modulates macrophage function. Ann N Y Acad Sci. 1070, 500-506. 2006

Herrera JL, Fernandez-Montesinos R, Gonzalez-Rey E, Delgado M, Pozo D. Protective role for plasmad DNA-mediated VIP gene transfer in non-obese diabetic mice. Ann N Y Acad Sci. 1070, 337-341. 2006

Chorny A, Gonzalez-Rey E, Fernandez-Martin A, Pozo D, Ganea D, Delgado M. Vasoactive intestinal peptide induces regulatory dendritic cells with therapeutic effects on autoimmune disorders. Proc Natl Acad Sci USA. 102, 13562-13567. 2005

C. Roodveldt, A. Aharoni, DS. Tawfik. Directed evolution of proteins for heterologous expression and stability. Current Opinion in Structural Biology. 15, 50-56. 2005.

A. Aharoni, L. Gaidukov, O. Khersonsky, S. McQ Gould, C. Roodveldt, DS. Tawfik. The 'evolvability' of promiscuous protein functions. Nature Genetics. 37, 73-76. 2005

Pozo D, Delgado M. The many faces of VIP in neuroimmunology: a cytokine rather a neuropeptide? FASEB Journal 18, 1325-1334. 2004

Delgado M, Pozo D, Ganea D. The significance of vasoactive intestinal peptide in immunomodulation. Pharmacological Reviews 56. 249-290. 2004

Pozo D. VIP- and PACAP-mediated immunomodulation as prospective therapeutic tools. Trends in Molecular Medicine 9, 211-217. 2003