Periodic Reporting for period 1 - TAONas-LUAD (Therapeutic Antisense Oligonucleotides Targeting NUMB Alternative Splicing in Lung Adenocarcinoma)
Berichtszeitraum: 2022-04-01 bis 2023-03-31
Lung adenocarcinomas (LUAD) represent 1/3 of all lung cancer cases. Despite notable advances, current treatments remain ineffective, resulting in <25% survival beyond 5 years. Due to the high heterogeneity of molecular abnormalities driving lung cancers, targeted therapies are applicable to only a small subset of patients. There is therefore an urgent unmet need for developing novel therapeutic approaches generally applicable to LUAD patients.
Alternative pre-mRNA splicing (AS) allows the synthesis of different protein variants from a single gene by differential selection of exonic sequences. Increased inclusion of exon 9 of the gene NUMB encodes a protein isoform that promotes cancer cell proliferation. This occurs in the vast majority of LUAD tumours, correlating with worse disease prognosis. Supported by the ERC PoC VALSL, we developed an innovative therapeutic approach based on the use of Antisense Oligonucleotides (AONs) that regulate NUMB AS. Our proprietary AONs correct NUMB pathological splicing, inhibit cancer cell proliferation and reduce tumour growth in four different mouse models of LUAD, including 2 Patient-Derived Xenograft models.
With support from the EIC Transition, we aim to bring this technology to a stage where it is ready to be validated in clinical trials. We will optimise our lead AONs by improving their chemistry, formulation and administration and will carry out regulatory pre-clinical studies. These will pave the way to the first application of AON-based splicing modulation in clinical oncology. To commercialise this technology, we also develop the business plan for a spin-off company, AON Therapeutics. In the long term, our project has the potential to generate compounds, presentations and delivery methods that can be applicable to other target AS events and/or cancer types, as well as to other AS-related diseases.
Alternative pre-mRNA splicing (AS) allows the synthesis of different protein variants from a single gene by differential selection of exonic sequences. Increased inclusion of exon 9 of the gene NUMB encodes a protein isoform that promotes cancer cell proliferation. This occurs in the vast majority of LUAD tumours, correlating with worse disease prognosis. Supported by the ERC PoC VALSL, we developed an innovative therapeutic approach based on the use of Antisense Oligonucleotides (AONs) that regulate NUMB AS. Our proprietary AONs correct NUMB pathological splicing, inhibit cancer cell proliferation and reduce tumour growth in four different mouse models of LUAD, including 2 Patient-Derived Xenograft models.
With support from the EIC Transition, we aim to bring this technology to a stage where it is ready to be validated in clinical trials. We will optimise our lead AONs by improving their chemistry, formulation and administration and will carry out regulatory pre-clinical studies. These will pave the way to the first application of AON-based splicing modulation in clinical oncology. To commercialise this technology, we also develop the business plan for a spin-off company, AON Therapeutics. In the long term, our project has the potential to generate compounds, presentations and delivery methods that can be applicable to other target AS events and/or cancer types, as well as to other AS-related diseases.
During RP1 (12 months), CRG identified a variety of AON sequences that efficiently promote exon skipping in vitro and compared five different chemical modifications as well as peptide conjugation, which led to the identification of optimized lead compounds.
Also, initial tests of various nanoparticle formulations identified combinations of lipid nanoparticles (LNP) and AON chemistries that compare favorably to lipofectamine-mediated cell transfection in vitro and can in principle allow different administration routes (intravenous, pulmonary) in vivo.
INSERM established a colony of the same NSG immune compromised mouse strain being used at CRG, which will accelerate progress and increase reproducibility of the results from lead compounds and formulations to assess their therapeutic effectiveness on NSG mice with A549 orthotopic xenografts. Also, to assess whether AONs promoting NUMB exon exclusion are more effective in cancer cells than in non-transformed cells, we took advantage of the bronchoalveolar human cell line BEAS-2B. BEAS-2B cell line is extensively used in lung studies as experimental model to investigate human airway epithelial cells. For instance, BEAS-2B have protective and regulatory mechanisms similar to primary human lung cells (PMID 7946385). Initial results strongly argue that modulation of NUMB exon inclusion is specific for transformed cells and therefore that targeting NUMB alternative splicing appears to be a special vulnerability of cancer cells, which is an important insight that further validates our efforts to develop a therapy based on splicing modulation of this gene.
Regarding IP and Regulatory activities, an Exploitation plan and a regulatory roadmap have been prepared by Inveniam and Asphalion, respectively. An Innovation Board (IB) has been set up and 1 dedicated IB-meeting was held.
Regarding Business Development and Venture Building, a Go-to-Market strategy and a preliminary Business Plan have been written, both important milestones for the launch of a spin-off company.
Also, initial tests of various nanoparticle formulations identified combinations of lipid nanoparticles (LNP) and AON chemistries that compare favorably to lipofectamine-mediated cell transfection in vitro and can in principle allow different administration routes (intravenous, pulmonary) in vivo.
INSERM established a colony of the same NSG immune compromised mouse strain being used at CRG, which will accelerate progress and increase reproducibility of the results from lead compounds and formulations to assess their therapeutic effectiveness on NSG mice with A549 orthotopic xenografts. Also, to assess whether AONs promoting NUMB exon exclusion are more effective in cancer cells than in non-transformed cells, we took advantage of the bronchoalveolar human cell line BEAS-2B. BEAS-2B cell line is extensively used in lung studies as experimental model to investigate human airway epithelial cells. For instance, BEAS-2B have protective and regulatory mechanisms similar to primary human lung cells (PMID 7946385). Initial results strongly argue that modulation of NUMB exon inclusion is specific for transformed cells and therefore that targeting NUMB alternative splicing appears to be a special vulnerability of cancer cells, which is an important insight that further validates our efforts to develop a therapy based on splicing modulation of this gene.
Regarding IP and Regulatory activities, an Exploitation plan and a regulatory roadmap have been prepared by Inveniam and Asphalion, respectively. An Innovation Board (IB) has been set up and 1 dedicated IB-meeting was held.
Regarding Business Development and Venture Building, a Go-to-Market strategy and a preliminary Business Plan have been written, both important milestones for the launch of a spin-off company.
Our project is contributing to the foundation of a new technology with potential high scientific impact. At the start of TAONas-LUAD, the technology was at a TRL3 level of maturity: we have demonstrated that antisense oligonucleotides (AONs) that modulate alternative splicing (AS) have anti-tumoral effects in mouse models of Lung Adenocarcinoma (LUAD), including Patient-Derived Xenografts. At the end of RP1, we are now progressing towards TRL4. The results are indeed encouraging, as the expansion of active chemical modifications and the effects of formulation into lipid nanoparticles with lung targeting properties give us hope that delivery of the optimized lead compounds in vivo will be improved and therefore that the already remarkable therapeutic effects obtained in mouse models of lung adenocarcinoma using naked AONs will be further improved to eventually generate a clinically useful drug.
In addition, Important insights about the mechanism of action of the AONs are being obtained, which could have implications in other cancer therapies.
In addition, Important insights about the mechanism of action of the AONs are being obtained, which could have implications in other cancer therapies.