Periodic Reporting for period 1 - EngiNano-HF (Engineered Nanoparticles Targeting Hair Follicles for Alopecia Areata Treatment)
Reporting period: 2022-06-01 to 2024-05-31
The EngiNano-HF project has focused on the treatment of Alopecia Areata, a common autoimmune disorder resulting from damage of the hair follicle by interferon-gamma (IFN-γ)-producing cytotoxic T lymphocytes and characterized by non-scarring hair loss. In most cases, hair falls out in small, round patches, leaving coin-sized areas of bare skin. Some estimates indicate that approximately 15 million people suffer from Alopecia Areata in Europe alone, and nearly 2% of the world population will develop this disease at some point during their lifetime. In 2015, Prof. Christiano et al. discovered that topically administered inhibitors of Janus kinases, a family of enzymes that are downstream effectors of IFN-γ cytokine receptors, prevent the development of Alopecia Areata and promote hair regrowth. Since this significant breakthrough, many pharmaceutical companies have been investing millions of euros in the development of novel Janus kinase inhibitors, several of them currently in phase 3 clinical trials, while Baricitinib was the first approved in June 2022 for the oral treatment of Alopecia Areata. Thus, it is expected that the global Alopecia Areata market size is poised to reach USD 1.6 billion by 2027. Therefore, the impact of EngiNano-HF is profound in both human and economic terms. Unfortunately, the oral delivery of Baricitinib displays severe side effects (for instance, vomits and headaches) that compromise the adhesion of the patients to the treatment. Under this scenario, the overall objective of EngiNano-HF is the design of nanoformulations for follicular delivery able to penetrate deeply into the hair follicle, which could dramatically improve the therapeutic outcome of Baricitinib while reducing side effects.
This MSCA Fellowship also aims to open the best career possibilities for Dr. Matías Picchio, a promising young scientist recently recognized by the Roal Society of Chemistry as an Emerging Investigator. After a successful PhD and 3 years of Post-Doc at CONICET (Argentina), he has decided to move to a new environment to
exploit his expertise in materials science fully. This multidisciplinary project has broadened the scientific knowledge of Dr. Picchio and helped him grow and expand his European and international networks to reach a future group leader position in the materials science field.
This MSCA Fellowship also aims to open the best career possibilities for Dr. Matías Picchio, a promising young scientist recently recognized by the Roal Society of Chemistry as an Emerging Investigator. After a successful PhD and 3 years of Post-Doc at CONICET (Argentina), he has decided to move to a new environment to
exploit his expertise in materials science fully. This multidisciplinary project has broadened the scientific knowledge of Dr. Picchio and helped him grow and expand his European and international networks to reach a future group leader position in the materials science field.
In the initial proposal of EngiNano-HF, thermoresponsive nanoparticles based on poly(vinyl caprolactam) (PVCL) and tannic acid (TA) as a supramolecular crosslinker were envisioned. These nanomaterials were prepared using a solvent-exchanged approach, where PVCL and TA were dissolved in ethanol as a hydrogen bond disrupting agent and then dropwise in water to establish TA crosslinking. Besides acting as a supramolecular crosslinker, TA has therapeutic properties as an antioxidant and anti-inflammatory, which is an additional advantage for Alopecia Areata, which is an inflammatory disorder. Different TA/PVCL mass ratios and surfactant (Tween 80) concentrations in the water phase were explored. In all cases, the obtained nanoparticles were unstable, leading to coagulation. Under this scenario, we activate our risk management plan and shift our nanoformulations to polydopamine nanoparticles (PDA NPs), another polyphenol with similar properties to TA. PDA NPs were then prepared by an oxidative process in ethanol/water mixtures using a strong base as an oxidizing agent (NH4OH). The reaction yield in all cases was around 50%. One attractive advantage of PDA NPs is that their size can be easily tuned by changing the NH4OH/dopamine molar ratio. We varied this parameter from 23.2 8.7 5.8 4.35 2.9 to 1.45 obtaining nanoparticles with sizes ranging from 150 to 950 nm, as determined by different characterization techniques. Some studies have reported that follicular penetration by nanoparticles is size-dependent. For this reason, we aimed to evaluate this parameter in a human skin ex-vivo model with variable follicle density. To do so, we loaded them with a hydrophobic dye, Nile red, to allow tracking by fluorescence. After dye loading and purification, we deposit the PDA NPs on excised human skin and incubate them for 24 h at 37°C. Skin from three different donors (2 female and 1 male) were used for the assays. After that, the skin was cut with a cryostat to obtain several segments of the hair follicles. Finally, confocal microscopy was used to image the penetration of the fluorescent dye. The obtained images are being processed with the help of the free software Image J. All these biological activities were performed during a 3-month secondment at the Charite Hospital (Berlin, Germany). For the next step, PDA NP showing the deepest follicular penetration will be used as a carrier of Baricitinib and submitted to new follicular penetration studies with human skin. During the project, we also advanced in a gel formulation for applying the PDA NPs on the scalp and helping their penetration. This gel formulation was based on eutectic technology using choline geranate and geranic acid (CAGE) with skin permeation ability. In the first stage, we evaluated the skin permeation of CAGE hosted in gelatin using non-encapsulated Nile red as a model hydrophobic molecule (like Baricitinib). The results showed that the eutectogel formulation could effectively assist the skin permeation of Nile red, and the next step will involve the inclusion of PDA NPs into these patches. Part of these results were recently published in Advanced Functional Materials (Impact Factor = 19), where Dr. Picchio as a corresponding author. Regarding dissemination, EngiNano-HF was promoted by Twitter (https://twitter.com/Polymat_BERC/status/1546404127516884992(opens in new window)) and the webpage of the project (https://matipicchio.wixsite.com/enginano-hf(opens in new window)) and their main results were presented in the outreach activity “11º Encuentro de Vidas Científicas” (https://twitter.com/eurekakutxa/status/1581922489109647360(opens in new window)) and as an invited speaker in the Biomaterials International (July 30-Aug. 3, 2023- Sapporo, Japon).
Polydopamine nanoparticles (PDA NPs) have not been explored for follicular delivery yet, and particularly their therapeutic properties as antioxidant and anti-inflammatory are beneficial for the Alopecia Areata treatment. Therefore, EngiNano-HF has introduced these robust nanocarriers and has gone beyond the state-of-the-art follicular penetration that can also serve for other inflammatory diseases as well. It is expected that the results of EngiNano-HF by the end of the project, demonstrating the effective follicular penetration of Nile red-loaded PDA NPs, open the way to advance to the loading and release studies of the Janus kinase inhibitor Baricitinib and their posterior evaluation in treating Alopecia Areata using hair follicle collapsed immune privilege ex-vivo models with the free drug as a control. These advances will have a profound socio-economic impact if the EngiNano-HF results can be validated with the recently FDA-approved Barcitinib as the first effective nanomedicine for treating Alopecia Areata. We count on the support of I+Med company, a partner of EngiNano-HF, to advance in a product formulation if the results with Baricitinib are successful. The societal implications of this achievement will represent a new dawn for patients suffering from Alopecia Areata, who often show depression, anxiety, and low self-esteem.