Periodic Reporting for period 2 - PAIN-Net (Molecule-to-man pain network)
Periodo di rendicontazione: 2019-02-01 al 2021-07-31
Neuropathic pain affects 5% of the general population and 40% of patients with neurological diseases, and has a key role in the pathophysiology of cancer pain that affects up to 50% of patients in the early disease stage and 30% of survivors, causing an enormous social burden. Treatments are inadequate with less than 50% of patients achieving 50% of pain relief at best, while up to 30% of cancer pain patients experience insufficient analgesia. Signatures of individual susceptibility to pain and analgesic responsiveness are urgently needed to improve patients’ management. Such advances are expected to originate from integrated clinical, basic science and entrepreneurial research readily translating scientific findings into benefits for patients. To consolidate these aims, a new generation of scientists with wide knowledge in neuropathic pain, focused research skills and experience in the interaction with biotechnology companies is needed. The PAIN-Net programme has supported such talented and inspired early stage researchers. Their research projects, embedded in an advanced molecule-to-man pain network, contributed to better understanding individual susceptibility to pain and analgesics responsiveness based on next generation sequencing, whole exome sequencing, epigenetics and pharmacogenomics studies, nociceptor and sodium channel functioning based on biophysics and proteomics studies, targeted analgesics based on high-throughput screening, and on the development and characterization of the first mouse models of sodium channel-related neuropathic pain.
ESR1 studied respectively: the validity of the Sudoscan device, the differences between evoked heat pain and spontaneous pain caused by small fiber neuropathy using functional-MRI and the accuracy of the skin biopsy technique by adding the proximal/distal ratio.
ESR2 has worked on a new way to diagnose neuropathic cancer pain, caused by damage to nerves due to the cancerous mass pressing effect and also identified factors influencing the way different patients respond to pain treatment.
ESR3 studied tiny nerve endings in human skin and their involvement in neuropathic pain. She discovered a difference in fine-tuning molecules between healthy and individuals suffering from chronic pain. Future experiments will show if, by modulation of identified molecules, we will be able to restore physiological pain perception.
ESR6 study aimed to identify genes and mutations responsible for neuropathic pain using next generation sequencing techniques. This approach revealed mutations in ion channel genes and novel gene candidates possibly involved in neuropathic pain, which were later tested in the lab setting for functional proof of pathogenicity.
ESR7 research project aimed to unravel a person's DNA in order to locate variants and determine its significance in association to pain. ESR7 selected variants not only in voltage-gated sodium channel genes, which are known to be responsible for neuropathic pain, but also in genes related to neurotransmission.
ESR8 observed that indeed painful and painless diabetic neuropathy patients have different DNA methylation patterns. In other words, it indicates that the experience of pain may not only results not from genetic mutations, but also be related to factors to which one is exposed during the life as diet, stress or physical activity, pollutants or toxins.
By measuring changes in the electrical pain signals of cells carrying mutations in sodium channels, ESR9 identified how they cause pain in patients as well as how they modulate the patients' responsiveness to certain drugs. These findings will help understanding how pain works and enable the development of personalized, more effective and better-tolerated treatments.
Using the Ion-Flux apparatus, ESR10 performed high-throughput screening of several drugs and found that Propranolol, Propafenone, Carvedilol and Nebivolol are inhibitors of TRPM8 ion channel. Such drugs could be considered as candidate for repurposing, or used as a lead compound for drug discovery.
ESR11 was able to replicate in mice the specific mutation in a sodium channel of patients suffering from pain. The mouse model was more sensitive to mechanical, heat and cold stimuli, similarly to patients.
ESR12 developed a mouse model using CRISPR-Cas9 technology. The characterization of this mouse model recapitulates human sodium channel-related painful neuropathy, which will be beneficial to explore novel drugs for painful neuropathy.
Chronic itch can make you scratch more and more and interfere with daily activities. To study this disease directly on human cells is difficult. ESR13 contributed to the production of the first mutant mouse model that showed similar disease symptom and could be helpful for future research to cure this disease in humans.
ESR14 (previously ESR4) studied the functional changes in the peripheral nervous structures of neuropathic pain patients and the replication of these phenomena in healthy people after irritating nociceptive afferents with physical and chemical methods (heat-capsaicin & ultraviolet radiation).
Using microneurography and the marking technique, ESR15 (previously ESR5) has been able to stimulate and record, for the first time ever, cold C-nociceptors. Also, Adelta fibers can be now recorded using this software, which can be used for clinical and preclinical trials to improve the diagnosis and drug development.
ESR2 has worked on a new way to diagnose neuropathic cancer pain, caused by damage to nerves due to the cancerous mass pressing effect and also identified factors influencing the way different patients respond to pain treatment.
ESR3 studied tiny nerve endings in human skin and their involvement in neuropathic pain. She discovered a difference in fine-tuning molecules between healthy and individuals suffering from chronic pain. Future experiments will show if, by modulation of identified molecules, we will be able to restore physiological pain perception.
ESR6 study aimed to identify genes and mutations responsible for neuropathic pain using next generation sequencing techniques. This approach revealed mutations in ion channel genes and novel gene candidates possibly involved in neuropathic pain, which were later tested in the lab setting for functional proof of pathogenicity.
ESR7 research project aimed to unravel a person's DNA in order to locate variants and determine its significance in association to pain. ESR7 selected variants not only in voltage-gated sodium channel genes, which are known to be responsible for neuropathic pain, but also in genes related to neurotransmission.
ESR8 observed that indeed painful and painless diabetic neuropathy patients have different DNA methylation patterns. In other words, it indicates that the experience of pain may not only results not from genetic mutations, but also be related to factors to which one is exposed during the life as diet, stress or physical activity, pollutants or toxins.
By measuring changes in the electrical pain signals of cells carrying mutations in sodium channels, ESR9 identified how they cause pain in patients as well as how they modulate the patients' responsiveness to certain drugs. These findings will help understanding how pain works and enable the development of personalized, more effective and better-tolerated treatments.
Using the Ion-Flux apparatus, ESR10 performed high-throughput screening of several drugs and found that Propranolol, Propafenone, Carvedilol and Nebivolol are inhibitors of TRPM8 ion channel. Such drugs could be considered as candidate for repurposing, or used as a lead compound for drug discovery.
ESR11 was able to replicate in mice the specific mutation in a sodium channel of patients suffering from pain. The mouse model was more sensitive to mechanical, heat and cold stimuli, similarly to patients.
ESR12 developed a mouse model using CRISPR-Cas9 technology. The characterization of this mouse model recapitulates human sodium channel-related painful neuropathy, which will be beneficial to explore novel drugs for painful neuropathy.
Chronic itch can make you scratch more and more and interfere with daily activities. To study this disease directly on human cells is difficult. ESR13 contributed to the production of the first mutant mouse model that showed similar disease symptom and could be helpful for future research to cure this disease in humans.
ESR14 (previously ESR4) studied the functional changes in the peripheral nervous structures of neuropathic pain patients and the replication of these phenomena in healthy people after irritating nociceptive afferents with physical and chemical methods (heat-capsaicin & ultraviolet radiation).
Using microneurography and the marking technique, ESR15 (previously ESR5) has been able to stimulate and record, for the first time ever, cold C-nociceptors. Also, Adelta fibers can be now recorded using this software, which can be used for clinical and preclinical trials to improve the diagnosis and drug development.
PAIN-Net offered a training programme combining academic and industrial expertise with theoretical knowledge and complementary skills, shaping a new generation of European neuroscientists fully equipped to explore the complexity of neuropathic pain. The local training-through- research that each ESRs received while conducting her/his individual project, has been integrated with a well-designed system of secondments, network-wide events and ESRs/group interactions. This granted a level of integration impossible to achieve in a national context, which has been further implemented by networking with neuroscience PhD courses of the Universities participating in the PAIN-Net consortium, thus fostering transfer of the PAIN-Net model into existing and/or new study plans. This innovated the structure of neuroscience ESR training in Europe, setting the ground for training collaborations that may extend beyond the funding period of this ETN.
In parallel, PAIN-Net research activities generated fundamental advances in the definition of individual susceptibility to pain and responsiveness to analgesics. The pathogenesis of pain in most patients is unknown and no biomarkers are available yet. Overall, not all individuals develop pain and it is currently not possible to predict who is more or less susceptible among those with similar risk exposure. Such variability remains unexplained yet. By performing research towards the identification of new therapeutic targets and strategies for pain treatment, while simultaneously training researchers to this end, the PAIN-Net project strengthened the European innovation capacity – with the aim of translating scientific findings in the complex field of pain into solutions able to benefit a larger percentage of patients - which is lagging behind.
In parallel, PAIN-Net research activities generated fundamental advances in the definition of individual susceptibility to pain and responsiveness to analgesics. The pathogenesis of pain in most patients is unknown and no biomarkers are available yet. Overall, not all individuals develop pain and it is currently not possible to predict who is more or less susceptible among those with similar risk exposure. Such variability remains unexplained yet. By performing research towards the identification of new therapeutic targets and strategies for pain treatment, while simultaneously training researchers to this end, the PAIN-Net project strengthened the European innovation capacity – with the aim of translating scientific findings in the complex field of pain into solutions able to benefit a larger percentage of patients - which is lagging behind.