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La presente invención se refiere una composición basada en biopolímeros que imita a la matriz extracelular natural del epitelio pigmentario de la retina (EPR) y que es útil para cultivo de células pluripotentes inducidas y del epitelio pigmentario de retina. La invención también se refiere al procedimiento de preparación de dicha composición y al uso de la misma.

Las enfermedades degenerativas de la retina (EDR) son un grupo heterogéneo de patologías, algunas hereditarias y otras con un origen complejo, que producen una pérdida progresiva de las células de la retina con la consiguiente pérdida de la agudeza visual pudiendo avanzar hasta producir una ceguera total. En los últimos años y gracias al desarrollo de terapias avanzadas como la terapia génica y celular muchos casos de ceguera podrían ser potencialmente tratables. El descubrimiento de las células madre de pluripotencia inducida o iPSC ha aportado una fuente celular ilimitada con gran potencial ya que gracias a sus propiedades son un instrumento prometedor para el estudio de enfermedades, así como para ser utilizadas en medicina regenerativa e ingeniería tisular. La edición génica es una novedosa tecnología con la se puede manipular con precisión una secuencia del genoma humano para lograr un efecto terapéutico mediante la cual se pueden corregir mutaciones que causan enfermedades, añadir genes terapéuticos a sitios específicos del genoma y eliminar genes letales o secuencias del genoma. En este trabajo se han generado y caracterizado con éxito dos líneas de iPSC a partir de muestras de sangre periférica de pacientes con EDR, como son la degeneración macular asociada a la edad (DMAE) y la retinosis pigmentaria (RP), con el fin de establecer modelos celulares de cada una de las enfermedades y terapia celular de reemplazo. Estas líneas nos han permitido conocer las bases celulares y moleculares que llevan al desarrollo de la enfermedad. Posteriormente, enfocándonos en la medicina regenerativa, con el objetivo de buscar nuevas terapias para tratar a pacientes con DMAE, las iPSC fueron diferenciadas hacia células de epitelio pigmentario de la retina (EPR). Las células obtenidas fueron caracterizadas y trasplantadas en diferentes modelos animales. Se demostrado que las células de EPR sobreviven y mantienen su fenotipo y orientación sin ningún efecto adverso local o sistémi

Mutations in the EYS gene are one of the major causes of autosomal recessive retinitis pigmentosa. EYS-retinopathy presents a severe clinical phenotype, and patients currently have no therapeutic options. The progress in personalised medicine and gene and cell therapies hold promise for treating this degenerative disease. However, lack of understanding and incomplete comprehension of disease's mechanism and the role of EYS in the healthy retina are critical limitations for the translation of current technical advances into real therapeutic possibilities. This review recapitulates the present knowledge about EYS-retinopathies, their clinical presentations and proposed genotype–phenotype correlations. Molecular details of the gene and the protein, mainly based on animal model data, are analysed. The proposed cellular localisation and roles of this large multi-domain protein are detailed. Future therapeutic approaches for EYS-retinopathies are discussed.

Retinal dystrophies associated to mutations in the CRB1 gene comprise a wide array of clinical presentations. A blood sample from a patient with a family history of CRB1-retinal dystrophy was used to prepare the iPSC line ESi082-A. The genotype of the donor, affected of a perifoveal-bilateral macular dystrophy includes one frameshift deletion and one hypomorphic allele. ESi082-A cell line has been characterized for pluripotency and will be used to prepare retinal cellular models to study the dysfunction leading to the disease.

The present invention relates to the therapeutic use of acylated piceid derivative compounds in ocular pathologies, in particular retinitis pigmentosa and in age-related macular degeneration.

Background Mutations in pre-mRNA splicing factor PRPF31 can lead to retinitis pigmentosa (RP). Although the exact disease mechanism remains unknown, it has been hypothesized that haploinsufficiency might be involved in the pathophysiology of the disease. Methods In this study, we have analyzed a mouse model containing the p.A216P mutation in Prpf31 gene. Results We found that mutant Prpf31 protein produces cytoplasmic aggregates in the retinal pigment epithelium and decreasing the protein levels of this splicing factor in the nucleus. Additionally, normal protein was recruited in insoluble aggregates when the mutant protein was overexpressed in vitro. In response to protein aggregation, Hspa4l is overexpressed. This member of the HSP70 family of chaperones might contribute to the correct folding and solubilization of the mutant protein, allowing its translocation to the nucleus. Conclusions Our data suggests that a mechanism haploinsufficiency and dominant-negative is involved in retinal degeneration due to mutations in PRPF31. HSP70 over-expression might be a new therapeutic target for the treatment of retinal degeneration due to PRPF31 mutations.

Damage to the retinal pigment epithelium (RPE) in age-related macular degeneration and other diseases results in photoreceptor cell death and blindness. Replacement of RPE is therefore being explored as a therapy for several retinal diseases. To move toward a future personalized autologous transplant approach, we have prepared a biocompatible implant using RPE derived from induced pluripotent stem cells (iPSCs) reprogrammed from a healthy donor's monocytes. The correct positioning of the polarized RPE is essential to fulfill its role and protect photoreceptors from degeneration. Hence, we have used a biocompatible hydrogel matrix of fibrin and agarose that allows the surgical placement of an RPE sheet in the subretinal space, keeping its functional orientation. Our aim was to demonstrate safety and viability of the transplant in preclinical models. Pigs were used to test the feasibility of a regular vitreoretinal surgery. Our results show that this implant is suitable for subretinal transplantation allowing human RPE cells to survive and maintain their phenotype and orientation without any local or systemic adverse events. The ability to transplant the iPSC-derived RPE sheet in its natural orientation will surely increase the chance to obtain a therapeutic effect in future translational studies. In the promising field of cellular therapy for retinal degenerative diseases, a new biomaterial is proposed as a scaffold to grow and surgically introduce a monolayer of retinal pigment epithelial cells into the subretinal space, keeping the orientation of the cells for a proper functional integration of the transplant. The use of induced pluripotent stem cells as the starting material for retinal pigment epithelial cells is intended to advance toward a personalized medicine approach.

Age-related macular degeneration (AMD) is the leading cause of adult blindness in developed countries and is characterized by progressive degeneration of the macula, the central region of the retina. A human induced pluripotent stem cell (hiPSC) line was derived from peripheral blood mononuclear cells (PBMCs) from a patient with a clinical diagnosis of dry AMD carrying the CFH Y402H polymorphism. Sendai virus was using for reprogramming and the pluripotent and differentiation capacity of the cells were assessed by immunocytochemistry and RT-PCR.

PRPF31 gene codes for a ubiquitously expressed splicing factor but mutations affect exclusively the retina, producing the progressive death of photoreceptor cells. We have identified a novel PRPF31 mutation in a patient with autosomal dominant retinitis pigmentosa. A blood sample was obtained and mononuclear cells were reprogrammed using the non-integrative Sendai virus to generate the cell line CABi001-A. The iPSC line has been characterized for pluripotency and differentiation capacity and will be differentiated toward photoreceptors and retinal pigment epithelium cells to study the molecular mechanism of the disease and test possible therapeutic strategies.

La presente invención se refiere al uso terapéutico de compuestos derivados piceido acilados en patologías oculares, en particular retinitis pigmentosa y en degeneración macular asociada a la edad, entre otras.

The present invention relates to the therapeutic use of acylated piceid derivative compounds in ocular pathologies, in particular retinitis pigmentosa and in age-related macular degeneration, inter alia. [EN], La presente invención se refiere al uso terapéutico de compuestos derivados piceido acilados en patologías oculares, en particular retinitis pigmentosa y en degeneración macular asociada a la edad, entre otras. [ES]

Retinitis pigmentosa (RP) is an inherited retinal degenerative disease. Mutations in EYS have been associated with autosomal recessive RP. The human iPS cell line, CABi002-A, derived from peripheral blood mononuclear cells from a patient carrying a heterozygous double mutation in EYS gene was generated by non-integrative reprogramming technology, using hOCT3/4, hSOX2, hc-MYC and hKLF4 reprogramming factors. Pluripotency and differentiation capacity were assessed by immunocytochemistry and RT-PCR. This iPSC line can be further differentiated towards the affected cells to understand the pathophysiology of the disease and test new therapeutic strategies.

[Aims]: Retinitis pigmentosa (RP) is an inherited disease characterized by a progressive degeneration of rod photoreceptors. An imbalance between pro- and antiapoptotic factors, such as Bax/Bcl-2, has been involved in retinal degeneration. To date, no cure or effective treatments are available for RP. Rasagiline is an antiparkinsonian drug that has shown neuroprotective effects in part attributed to a modulation of Bax/Bcl-2 expression. In this study, we have evaluated the use of rasagiline as a potential treatment for RP. [Methods]: Newborn rd10 mice, a RP model, were treated with oral rasagiline during 30 days followed by a functional and morphological characterization of their mouse retinas. [Results]: Treated animals showed a significant improvement in visual acuity and in the electrical responses of photoreceptors to light stimuli. Rasagiline delayed photoreceptor degeneration, which was confirmed not only by a high photoreceptor nuclei counting, but also by a sustained expression of photoreceptor-specific markers. In addition, the expression of proapoptotic Bax decreased, whereas the antiapoptotic factor Bcl-2 increased after rasagiline treatment. [Conclusion]: This study provides new evidences regarding the neuroprotective effect of rasagiline in the retina, and it brings new insight into the development of future clinical trials using this well-established antiparkinsonian drug to treat RP.

Patient-derived cellular models can provide a specific tool to test new therapeutic approaches for retinal degenerative diseases. Mutations in AIPL1 are associated with Leber Congenital Amaurosis (LCA). Considering that AIPL1 protein is only expressed in photoreceptors we decided to establish a cellular model of AIPL1-LCA by differentiation of induced pluripotent stem cells (iPS) towards photoreceptor lineage.

It has been report ed that Sirt1 nuclear expression decreases in the outer nuclear layer (ONL) of retinal degeneration 10 (rd10 ) mice at postnatal day 15 (P15), and this abnormal pattern is correlated with the beginning of photoreceptor degeneration. Sirt1 is a class III histone deacetylase, and over-­expression of these p roteins have shown neuroprotective effect in several experimental models of neurodegenerative diseases. However, it is still unknown whether Sirt1 modulation could induce neuroprotection of ph otoreceptors. We tested the hypothesis that subretinal injection of new Sirt1 activators would slow degeneration of photoreceptors using an experimental mouse model of autosomal recessive retinitis pigmentosa (RP).

Retinitis pigmentosa (RP) is the most common cause of inherited blindness in adults. Mutations in the PRPF31 gene produce autosomal dominant RP (adRP). To date there are no effective treatments for this disease. The purpose of this study was to design an efficient non-viral vector for human PRPF31 gene delivery as an approach to treat this form of adRP. Span based nanoparticles were developed to mediate gene transfer in the subretinal space of a mouse model of adRP carrying a point mutation (A216P) in the Prpf31 gene. Funduscopic examination, electroretinogram, optomotor test and optical coherence tomography were conducted to further in vivo evaluate the safety and efficacy of the nanosystems developed. Span-polyarginine (SP-PA) nanoparticles were able to efficiently transfect the GFP and PRPF31 plasmid in mice retinas. Statistically significant improvement in visual acuity and retinal thickness were found in Prpf31 mice treated with the SP-PA-PRPF31 nanomedicine.