Periodic Reporting for period 1 - Pulmonary Fibrosis (Glycomic and Genomic Repercussion of Nebulisable Gal-3 Inhibitory Medical Device Treatment in Pulmonary Fibrosis)
Période du rapport: 2022-09-01 au 2024-08-31
Idiopathic pulmonary fibrosis (IPF) is the most common of idiopathic interstitial pneumonia with an increasing prevalence due to aged population, lifestyle and environmental exposure [1]. Today there is no cure for IPF, there is only two approved treatments that slow down progression. There is a need for new and more efficient anti-fibrotic treatments that stop and resolve fibrosis in IPF. In the current project, we were exploring the conjugation of a novel anti-fibrotic drug (TD139) to macromolecular polyethylene glycol (PEG) through a cleavable link to improve nebulization delivery, alveolar retention and prolonged delivery of TD139. Unfortunately, TD139 was discontinued from Phase II clinical trials do to worsening of effects [2]. We are exploring another anti-fibrotic drug, sobetirome, which the secondment institution has experimental experience [3]. The use of biomaterial delivery strategies has the potential to increase efficiency of anti-fibrotic drugs by increasing bioactive time of action and patient compliance.
The objectives are:
• Conjugate TD139 to PEG through a cleavable bond.
• Achieve prolonged physiological TD139 deliveries maintaining biological action.
• Increased TD139 retention in the lung alveoli.
• Achieve efficient nebulization of PEG-TD139 conjugates.
• Optimize in vitro and in vivo models for anti-fibrotic test of TD139 delivery.
• Compare in vivo treatment performance of PEG-TD139 to gold standard Nintedanib.
• Obtain human data using human precision cut lung slices.
[1] s12325-022-02395-9; [2] NCT03832946, [3] 10.1038/nm.4447
The objectives are:
• Conjugate TD139 to PEG through a cleavable bond.
• Achieve prolonged physiological TD139 deliveries maintaining biological action.
• Increased TD139 retention in the lung alveoli.
• Achieve efficient nebulization of PEG-TD139 conjugates.
• Optimize in vitro and in vivo models for anti-fibrotic test of TD139 delivery.
• Compare in vivo treatment performance of PEG-TD139 to gold standard Nintedanib.
• Obtain human data using human precision cut lung slices.
[1] s12325-022-02395-9; [2] NCT03832946, [3] 10.1038/nm.4447
We have optimized and validated the covalent conjugation of TD139 to PEG with two cleavable moieties that allow spontaneous release of TD139 in its original form ranging from a few days to a couple of months under physiological conditions. The PEG-TD139 conjugates are soluble and can be nebulized.
We have tested the anti-fibrotic effect of TD139 in different in vitro, mice and human precision cut lung slices and mice in vivo models to validate the efficiency of the conjugates. Unfortunately, any of these models have shown a remarkable anti-fibrotic effect which is in line with the current discontinuation of TD139 from clinical trials. We are exploring the conjugation of sobetirome, another novel anti-fibrotic drug, which anti-fibrotic effects have been tested in the secondment institution.
Parallelly, I have been involved in the validation of Raman spectroscopy for the diagnosis of IPF among patients with interstitial lung disease (ILD). The diagnosis of IPF is complex, unprecise and time-consuming. Raman spectroscopy is sensitive to molecular changes and can be used to classify different tissue states [4]. Using big data processing and discriminant analysis we have been able to predict IPF and non-IPF outcome in a small cohort of IPF and non-IPF ILD samples with a decent accuracy. We are working in the inclusion of more measurements to improve the accuracy of the analysis.
I am also working in the validation of a variation of the precision cut lung slice (PCLS) method used in my secondment institution. PCLS allows for ex-vivo study of animal and human lungs [5]. This reduces animal intervention and allows obtaining human data. The current method uses agarose lung inflation for vibratome cutting. The agarose maintains lung structure; however, it significantly reduces yields of RNA extraction and maintains the lung in an artificial embedded state that we belive hinders cell communication. We know by single-cell RNA sequencing that there is a significant change in gene expression during culture conditions. However, there are not drastic histological changes. By inflating the lungs with gelatin, this embedded state is removed (gelatin dissolves at 37 °C). We have observed that in the absence of a gel matrix there is a quick lung remodeling response developing in fibrosis. RNA yields and house keeping expression are orders of magnitude higher than with conventional agarose-PCLS. This model has potential to study aberrant cell populations and initial lung abnormalities that lead to fibrosis. We are validating this model in mouse and human samples.
[4] j.actbio.2023.03.016 [5] ajplung.00084.2017
We have tested the anti-fibrotic effect of TD139 in different in vitro, mice and human precision cut lung slices and mice in vivo models to validate the efficiency of the conjugates. Unfortunately, any of these models have shown a remarkable anti-fibrotic effect which is in line with the current discontinuation of TD139 from clinical trials. We are exploring the conjugation of sobetirome, another novel anti-fibrotic drug, which anti-fibrotic effects have been tested in the secondment institution.
Parallelly, I have been involved in the validation of Raman spectroscopy for the diagnosis of IPF among patients with interstitial lung disease (ILD). The diagnosis of IPF is complex, unprecise and time-consuming. Raman spectroscopy is sensitive to molecular changes and can be used to classify different tissue states [4]. Using big data processing and discriminant analysis we have been able to predict IPF and non-IPF outcome in a small cohort of IPF and non-IPF ILD samples with a decent accuracy. We are working in the inclusion of more measurements to improve the accuracy of the analysis.
I am also working in the validation of a variation of the precision cut lung slice (PCLS) method used in my secondment institution. PCLS allows for ex-vivo study of animal and human lungs [5]. This reduces animal intervention and allows obtaining human data. The current method uses agarose lung inflation for vibratome cutting. The agarose maintains lung structure; however, it significantly reduces yields of RNA extraction and maintains the lung in an artificial embedded state that we belive hinders cell communication. We know by single-cell RNA sequencing that there is a significant change in gene expression during culture conditions. However, there are not drastic histological changes. By inflating the lungs with gelatin, this embedded state is removed (gelatin dissolves at 37 °C). We have observed that in the absence of a gel matrix there is a quick lung remodeling response developing in fibrosis. RNA yields and house keeping expression are orders of magnitude higher than with conventional agarose-PCLS. This model has potential to study aberrant cell populations and initial lung abnormalities that lead to fibrosis. We are validating this model in mouse and human samples.
[4] j.actbio.2023.03.016 [5] ajplung.00084.2017
We expect to conjugate sobetirome to PEG and test its efficiency in vitro, ex-vivo mice and human PCLS and in vivo mouse. We expect to publish the conjugation of TD139 or sobetirome in a lung biology or biomaterials journal such as European Respiratory Journal or Advance Materials. We plan to present the results in the next European Respiratory Society (ERS) Congress in Amsterdam. We will study patentability with the involved institutions. The marketing of more efficient anti-fibrotic delivery strategies has the potential to improve treatment outcomes in IPF patients and reduce hospitalization costs. It is estimated a healthcare cost of $2 billion for IPF annually in the US [6].
We also plan to increase the number of measurements in the Raman characterization of IPF samples to reinforce and improve the diagnostic accuracy. We expect to publish these findings in a diagnostic journal such as Theranostics. We also plan to present result outcomes in the next European Respiratory Society (ERS) Congress. We will also study patentability of the developed methodology. Automated and precise diagnostic tools for IPF have the potential for an earlier treatment implementation reducing both disease-related and diagnostics costs. A similar methodology can be implemented for the biopsy diagnosis of other ILD subgroups or pathologies in other organs.
We expect to validate the gelatin-PCLS model using single-cell RNAseq and selected protein markers to identify aberrant cell populations leading to fibrosis. The study of aberrant cells leading to lung fibrosis have the potential to unveil causing mechanisms, earlier diagnostic markers and new treatment targets. I plan to use the generated preliminary data to apply for my next research grant such as Starting Grant European Research Council (ERC).
[6] AnnalsATS.201412-553OC
We also plan to increase the number of measurements in the Raman characterization of IPF samples to reinforce and improve the diagnostic accuracy. We expect to publish these findings in a diagnostic journal such as Theranostics. We also plan to present result outcomes in the next European Respiratory Society (ERS) Congress. We will also study patentability of the developed methodology. Automated and precise diagnostic tools for IPF have the potential for an earlier treatment implementation reducing both disease-related and diagnostics costs. A similar methodology can be implemented for the biopsy diagnosis of other ILD subgroups or pathologies in other organs.
We expect to validate the gelatin-PCLS model using single-cell RNAseq and selected protein markers to identify aberrant cell populations leading to fibrosis. The study of aberrant cells leading to lung fibrosis have the potential to unveil causing mechanisms, earlier diagnostic markers and new treatment targets. I plan to use the generated preliminary data to apply for my next research grant such as Starting Grant European Research Council (ERC).
[6] AnnalsATS.201412-553OC