Periodic Reporting for period 4 - FutureTrophicFactors (Elucidating therapeutic effects and mode of action of future trophic factorsin ALS and Parkinson’s disease)
Periodo di rendicontazione: 2023-08-01 al 2024-01-31
The prevalence of neurodegenerative diseases such as Parkinson’s disease (PD) and amyotrophic lateral sclerosis (ALS) is growing rapidly due to an aging population and increased life
expectancy. Current treatments for PD and ALS only relieve symptoms and cannot stop the progression of the disease, thus there is an urgent need for new therapies.
Neurotrophic factors (NTFs) are secretory proteins that regulate the development, function and survival of neurons. They have been explored as novel drugs for the treatment of PD and ALS but their efficacy in clinical
trials has been modest.
Cerebral dopamine neurotrophic factor (CDNF) is a protein with NTF properties. I have shown that it protects and restores the function of dopamine (DA) neurons in rodent toxin models of PD more
effectively than other NTFs making it promising drug candidates for the disease-modifying treatment of PD. CDNF showed neurorestorative effects in non-human primate models of PD. In addition, CDNF was safe and successfully passed phase 1/2
clinical trials on PD patients. My group has also shown that CDNF improves motor coordination and protects motoneurons (MNs) in three different
genetic animal model of ALS (De Lorenzo et al., Brain)
Despite encouraging results with CDNF in animal models and clinical trials of PD and ALS, protein-based treatments have drawbacks, requiring direct delivery to the brain through invasive surgery, since
they cannot pass through the blood brain barrier (BBB). My recent discovery, however, may overcome this difficulty: novel CDNF variant protects DA and motoneurons in
vitro and in vivo and can pass through the BBB. Importantly, we have found that subcutaneous administration of the novel CDNF fragment improves motor function and is
neuroprotective in animal models of PD and ALS.
This is important for the society, bacause an aging population and increased life expectancy in developed countries are leading to a higher incidence of age-related neurodegenerative diseases such as ALS and PD.
Treating these diseases is a growing economic burden for health care systems worldwide. In Europe, the estimated total cost of brain disorders is €798 billion per year in 2010 prices (including direct costs of treatment and care plus indirect cost of lost workdays and
lost productivity). ALS and PD are incurable conditions and eventually lead to death. There is thus a huge unmet need for new and more effective treatments that could slow down or even stop disease progression.
Neurodegenerative diseases are characterized by the vulnerability of specific neural populations, giving rise to a specific set of symptoms. Nevertheless, pathogenesis of neurodegenerative diseases, such as ALS and PD,
shows striking similarities, suggesting, that discoveries in one condition may benefit others.
Objectives were:
Objective 1) Study and Compare the Mode of Action of CDNF and CDNF fragment, 2) study the effect of Systemic Delivery of BBB penetrating CDNF fragment in Neurodegenerative Diseases and 3) Developing Human MNs and DA Neurons from Patient-derived iPS cells and Testing the Effects of CDNF fragments on Survival, ER stress and Reduction of Protein Aggregates in these Cells
expectancy. Current treatments for PD and ALS only relieve symptoms and cannot stop the progression of the disease, thus there is an urgent need for new therapies.
Neurotrophic factors (NTFs) are secretory proteins that regulate the development, function and survival of neurons. They have been explored as novel drugs for the treatment of PD and ALS but their efficacy in clinical
trials has been modest.
Cerebral dopamine neurotrophic factor (CDNF) is a protein with NTF properties. I have shown that it protects and restores the function of dopamine (DA) neurons in rodent toxin models of PD more
effectively than other NTFs making it promising drug candidates for the disease-modifying treatment of PD. CDNF showed neurorestorative effects in non-human primate models of PD. In addition, CDNF was safe and successfully passed phase 1/2
clinical trials on PD patients. My group has also shown that CDNF improves motor coordination and protects motoneurons (MNs) in three different
genetic animal model of ALS (De Lorenzo et al., Brain)
Despite encouraging results with CDNF in animal models and clinical trials of PD and ALS, protein-based treatments have drawbacks, requiring direct delivery to the brain through invasive surgery, since
they cannot pass through the blood brain barrier (BBB). My recent discovery, however, may overcome this difficulty: novel CDNF variant protects DA and motoneurons in
vitro and in vivo and can pass through the BBB. Importantly, we have found that subcutaneous administration of the novel CDNF fragment improves motor function and is
neuroprotective in animal models of PD and ALS.
This is important for the society, bacause an aging population and increased life expectancy in developed countries are leading to a higher incidence of age-related neurodegenerative diseases such as ALS and PD.
Treating these diseases is a growing economic burden for health care systems worldwide. In Europe, the estimated total cost of brain disorders is €798 billion per year in 2010 prices (including direct costs of treatment and care plus indirect cost of lost workdays and
lost productivity). ALS and PD are incurable conditions and eventually lead to death. There is thus a huge unmet need for new and more effective treatments that could slow down or even stop disease progression.
Neurodegenerative diseases are characterized by the vulnerability of specific neural populations, giving rise to a specific set of symptoms. Nevertheless, pathogenesis of neurodegenerative diseases, such as ALS and PD,
shows striking similarities, suggesting, that discoveries in one condition may benefit others.
Objectives were:
Objective 1) Study and Compare the Mode of Action of CDNF and CDNF fragment, 2) study the effect of Systemic Delivery of BBB penetrating CDNF fragment in Neurodegenerative Diseases and 3) Developing Human MNs and DA Neurons from Patient-derived iPS cells and Testing the Effects of CDNF fragments on Survival, ER stress and Reduction of Protein Aggregates in these Cells
In the beginning of the ERC project we investigated whether CDNF had a neuroprotective effect in vivo in the SOD1-G93A mouse, an ALS in vivo animal model where chronic ER stress response in the corresponding MNs has been previously reported in many studies. To this end, we utilized a single injection of CDNF, which in PD animal models effectively counteracted dopamine neuron degeneration [Lindholm et al., 2007]. As proof-of principle, we first validated that CDNF and 125I-labelled CDNF injected in the brain lateral ventricle efficiently diffuse to different areas of the brain, including cortex, striatum, and substantia nigra, and to the lumbar spinal cord. Moreover, we found that CDNF specifically co-localizes with lumbar MNs. Early symptomatic SOD1-G93A mice and WT littermates of 13 weeks of age received a single i.c.v. injection of 10 μg of human CDNF or phosphate buffered saline (PBS) as vehicle and were examined twice a week to follow changes in symptoms and motor behavior until the final stage of paralysis. At the time of treatment, SOD1-G93A mice displayed measurable tremors in the hind limbs. Upon a single CDNF injection, mutant mice developed gait impairment and paralysis symptoms significantly more slowly than PBS-treated mutant littermates, and survival was increased in both female and male SOD1 mice. Along with this, their balance and motor behavior performance were significantly ameliorated by CDNF treatment. One week after CDNF or PBS injection, the CDNF group showed an increased ability to run on the smallest 8 mm rod in the multiple static rods experimental paradigm for females and on the 21 and 11 mm rods for males, compared to the PBS group. These gender differences in the experimental tasks are in accordance with previous observations that males develop major symptoms approximately one week earlier than female littermates. In the accelerating rotarod, CDNF-treated mice showed an increased latency to fall compared to vehicle treated-mice. No statistical difference was found in CDNF or vehicle-injected WT littermates over time.
In the open field, SOD1-G93A mice exhibited a decreased number of rearings compared to WT mice, which is associated with less strength in the hind limb muscles, and CDNF treatment increased the number of rearings compared to PBS controls at 16 weeks. To correlate the improvement of motor behavior to MN survival, we examined the lumbar spinal cord, which revealed a significantly higher number of MNs present in the CDNF-treated compared to PBS-treated mice. The results showing that intracerebroventricularly injected CDNF protects spinal motoneurons in 3 different transgenic rodent models of ALS were published in Brain 2023.
In the ERC StG project we tested the effect of CDNF and CDNF fragment in continuous cells, primary Motoneuron and DA neuron cultures and in several animal models of ALS (SOD1 and TDP-43 rodent ) and PD (6-OHDA and MPTP toxin models ).
We were able to see efficacy after single or chronic intracerebroventraicular administration of CDNF and CDNF fragment in SOD1-G93A mouse model of ALS. We were also able to show neuroprotection after CDNF and CDNF fragment icv infusion in TDP-43 rat model of ALS, in which no other research group has ever reported positive effects with drug candidates. The experiments were carried out as planned
In the open field, SOD1-G93A mice exhibited a decreased number of rearings compared to WT mice, which is associated with less strength in the hind limb muscles, and CDNF treatment increased the number of rearings compared to PBS controls at 16 weeks. To correlate the improvement of motor behavior to MN survival, we examined the lumbar spinal cord, which revealed a significantly higher number of MNs present in the CDNF-treated compared to PBS-treated mice. The results showing that intracerebroventricularly injected CDNF protects spinal motoneurons in 3 different transgenic rodent models of ALS were published in Brain 2023.
In the ERC StG project we tested the effect of CDNF and CDNF fragment in continuous cells, primary Motoneuron and DA neuron cultures and in several animal models of ALS (SOD1 and TDP-43 rodent ) and PD (6-OHDA and MPTP toxin models ).
We were able to see efficacy after single or chronic intracerebroventraicular administration of CDNF and CDNF fragment in SOD1-G93A mouse model of ALS. We were also able to show neuroprotection after CDNF and CDNF fragment icv infusion in TDP-43 rat model of ALS, in which no other research group has ever reported positive effects with drug candidates. The experiments were carried out as planned
We have successfully compared the effects of CDNF and CDNF fragment in human motoneurons. We have been also able to select the correct dose and tested the effects of subcutaneously injected CDNF fragment and tested the effects of CDNF fragment in rodents models of PD and ALS
We have prepared the first manuscript ( characterization of CDNF fragment) that will be submitted to Bioarchives Spring 2024
We have prepared the first manuscript ( characterization of CDNF fragment) that will be submitted to Bioarchives Spring 2024