THOMAZ ANTONIO FLEURY CURADO
Projetos de Pesquisa
Unidades Organizacionais
LIM/63, Hospital das Clínicas, Faculdade de Medicina
8 resultados
Resultados de Busca
Agora exibindo 1 - 8 de 8
- Designer Receptors Exclusively Activated by Designer Drugs Approach to Treatment of Sleep-disordered Breathing(2021) CURADO, Thomaz Fleury; PHO, Huy; FREIRE, Carla; AMORIM, Mateus R.; BONAVENTURA, Jordi; KIM, Lenise J.; LEE, Rachel; CABASSA, Meaghan E.; STREETER, Stone R.; BRANCO, Luiz G.; SENNES, Luiz U.; FISHBEIN, Kenneth; SPENCER, Richard G.; SCHWARTZ, Alan R.; BRENNICK, Michael J.; MICHAELIDES, Michael; FULLER, David D.; POLOTSKY, Vsevolod Y.Rationale: Obstructive sleep apnea is recurrent upper airway obstruction caused by a loss of upper airway muscle tone during sleep. The main goal of our study was to determine if designer receptors exclusively activated by designer drugs (DREADD) could be used to activate the genioglossus muscle as a potential novel treatment strategy for sleep apnea. We have previously shown that the prototypical DREADD ligand clozapine-N-oxide increased pharyngeal diameter in mice expressing DREADD in the hypoglossal nucleus. However, the need for direct brainstem viral injections and clozapine-N-oxide toxicity diminished translational potential of this approach, and breathing during sleep was not examined. Objectives: Here, we took advantage of our model of sleep-disordered breathing in diet-induced obese mice, retrograde properties of the adeno-associated virus serotype 9 (AAV9) viral vector, and the novel DREADD ligand J60. Methods: We administered AAV9-hSyn-hM3(Gq)-mCherry or control AAV9 into the genioglossus muscle of diet-induced obese mice and examined the effect of J60 on genioglossus activity, pharyngeal patency, and breathing during sleep. Measurements and Main Results: Compared with control, J60 increased genioglossus tonic activity by greater than sixfold and tongue uptake of 2-deoxy-2-[F-18]fluoro-D-glucose by 1.5-fold. J60 increased pharyngeal patency and relieved upper airway obstruction during non-REM sleep. Conclusions: We conclude that following intralingual administration of AAV9-DREADD, J60 can activate the genioglossus muscle and improve pharyngeal patency and breathing during sleep.
- Neurostimulation Treatment of OSA(2018) CURADO, Thomaz Fleury; OLIVEN, Arie; SENNES, Luiz U.; POLOTSKY, Vsevolod Y.; EISELE, David; SCHWARTZ, Alan R.Over the past 30 years, hypoglossal nerve stimulation has moved through a development pathway to become a viable treatment modality for patients with OSA. Initial pilot studies in animals and humans laid the conceptual foundation for this approach, leading to the development of fully implantable stimulating systems for therapeutic purposes. These devices were then shown to be both safe and efficacious in feasibility studies. One such closed-loop stimulating device was found to be effective in treating a limited spectrum of apneic patients and is currently approved by the US Food and Drug Administration for this purpose. Another open-loop stimulating system is currently being rigorously tested in a pivotal trial. Collectively, clinical trials of hypoglossal nerve stimulating systems have yielded important insights that can help optimize therapeutic responses to hypoglossal nerve stimulation. These insights include specific patient selection criteria and methods for delivering stimulation to specific portions of the hypoglossal nerve and/or genioglossus muscle. New approaches for activating efferent and afferent motor pathways are currently in early-stage laboratory development and hold some long-term promise as a novel therapy.
- Intranasal Leptin Prevents Opioid-induced Deaths in Mice(2021) FREIRE, Carla; PHO, Huy; FONTI, Shannon; KIM, Lenise; FLEURY-CURADO, Thomaz; SENNES, Luiz; POLOTSKY, Vsevolod
- Sleep-disordered breathing in C57BL/6J mice with diet-induced obesity(2018) CURADO, Thomaz Fleury; PHO, Huy; BERGER, Slava; CABALLERO-ERASO, Candela; SHIN, Mi-Kyung; SENNES, Luiz Ubirajara; Luu Pham; SCHWARTZ, Alan R.; POLOTSKY, Vsevolod Y.Obesity leads to sleep-disordered breathing (SDB) manifested by recurrent upper airway obstructions termed obstructive sleep apnea (OSA) and carbon dioxide retention due to hypoventilation. The objective of this work was to characterize breathing during sleep in C57BL6/J mice with diet-induced obesity (DIO). Arterial blood gas was measured in nine obese and nine lean mice during wakefulness. Nine male mice with DIO and six lean male C57BL/6J mice were head mounted with electroencephalogram (EEG) and electromyogram (EMG) electrodes. Sleep recordings were performed in the whole body plethysmography chamber; upper airway obstruction was characterized by the presence of inspiratory flow limitation in which airflow plateaus with increases in inspiratory effort. Obese mice showed significantly lower pH and higher partial pressure of arterial CO2 (PaCO2) in arterial blood gas compared to lean mice, 7.35 +/- 0.04 versus 7.46 +/- 0.06 (p < 0.001) and 38 +/- 8 mm Hg versus 30 +/- 5 mm Hg (p < 0.001). Obese mice had similar levels of minute ventilation to lean mice during sleep and wakefulness, despite higher body weight and temperature, indicating an increase in the metabolic rate and hypoventilation. Obese mice also showed baseline hypoxemia with decreased mean oxyhemoglobin saturation across sleep/wake states. Obese mice had a higher prevalence of flow-limited breathing compared to lean mice during sleep. However, the oxygen desaturation index in lean and obese mice did not differ. We conclude that DIO in mice leads to hypoventilation. Obesity also increases the frequency of inspiratory limited breaths, but it does not translate into progression of OSA.
- Silencing of Hypoglossal Motoneurons Leads to Sleep Disordered Breathing in Lean Mice(2018) CURADO, Thomaz A. Fleury; PHO, Huy; DERGACHEVA, Olga; BERGER, Slava; LEE, Rachel; FREIRE, Carla; ASHEROV, Aya; SENNES, Luis U.; MENDELOWITZ, David; SCHWARTZ, Alan R.; POLOTSKY, Vsevolod Y.Obstructive Sleep Apnea (OSA) is a prevalent condition and a major cause of morbidity and mortality in Western Society. The loss of motor input to the tongue and specifically to the genioglossus muscle during sleep is associated with pharyngeal collapsibility and the development of OSA. We applied a novel chemogenetic method to develop a mouse model of sleep disordered breathing Our goal was to reversibly silence neuromotor input to the genioglossal muscle using an adeno-associated viral vector carrying inhibitory designer receptors exclusively activated by designer drugs AAV5-hM4Di-mCherry (DREADD), which was delivered bilaterally to the hypoglossal nucleus in fifteen C57BU6J mice. In the in vivo experiment, 4 weeks after the viral administration mice were injected with a DREADD ligand clozapine-N-oxide (CNO, i.p., 1mg/kg) or saline followed by a sleep study; a week later treatments were alternated and a second sleep study was performed. Inspiratory flow limitation was recognized by the presence of a plateau in mid-respiratory flow; oxyhemoglobin desaturations were defined as desaturations > 4% from baseline. In the in vitro electrophysiology experiment, four males and three females of 5 days of age were used. Sixteen-nineteen days after DREADD injection brain slices of medulla were prepared and individual hypoglossal motoneurons were recorded before and after CNO application. Positive mCherry staining was detected in the hypoglossal nucleus in all mice confirming successful targeting. In sleep studies, CNO markedly increased the frequency of flow limitation n NREM sleep (from 1.9 +/- 1.3% after vehicle injection to 14.2 +/- 3.4% after CNO, p < 0.05) and REM sleep (from 22.3% +/- 4.1% to 30.9 +/- 4.6%, respectively, p < 0.05) compared to saline treatment, but there was no significant oxyhemoglobin desaturation or sleep fragmentation. Electrophysiology recording in brain slices showed that CNO inhibited firing frequency of DREADD-containing hypoglossal motoneurons. We conclude that chemogenetic approach allows to silence hypoglossal motoneurons in mice, which leads to sleep disordered breathing manifested by inspiratory flow limitation during NREM and REM sleep without oxyhemoglobin desaturation or sleep fragmentation. Other co-morbid factors, such as compromised upper airway anatomy, may be needed to achieve recurrent pharyngeal obstruction observed in OSA.
- Intranasal Leptin Prevents Opioid-induced Sleep-disordered Breathing in Obese Mice(2020) FREIRE, Carla; PHO, Huy; KIM, Lenise J.; WANG, Xin; DYAVANAPALLI, Jhansi; STREETER, Stone R.; FLEURY-CURADO, Thomaz; SENNES, Luiz U.; MENDELOWITZ, David; POLOTSKY, Vsevolod Y.Respiratory depression is the main cause of morbidity and mortality associated with opioids. Obesity increases opioid-related mortality, which is mostly related to comorbid obstructive sleep apnea. Naloxone, a mu-opioid receptor blocker, is an effective antidote, but it reverses analgesia. Like humans with obesity, mice with diet-induced obesity hypoventilate during sleep and develop obstructive sleep apnea, which can be treated with intranasal leptin. We hypothesized that intranasal leptin reverses opioid-induced sleep-disordered breathing in obese mice without decreasing analgesia. To test this hypothesis, mice with diet-induced obesity were treated with morphine at 10 mg/kg subcutaneously and with leptin or placebo intranasally. Sleep and breathing were recorded by barometric plethysmography, and pain sensitivity was measured by the tail-flick test. Excitatory postsynaptic currents were recorded in vitro from hypoglossal motor neurons after the application of the mu-opioid receptor agonist [D-Ala(2), N-MePhe(4), Gly-ol]-enkephalin and leptin. Morphine dramatically increased the frequency of apneas and greatly increased the severity of hypoventilation and obstructive sleep apnea. Leptin decreased the frequency of apneas, improved obstructive sleep apnea, and completely reversed hypoventilation, whereas morphine analgesia was enhanced. Our in vitro studies demonstrated that [D-Ala(2), N-MePhe(4), Gly-ol]-enkephalin reduced the frequency of excitatory postsynaptic currents in hypoglossal motoneurons and that application of leptin restored excitatory synaptic neurotransmission. Our findings suggest that intranasal leptin may prevent opioid respiratory depression during sleep in patients with obesity receiving opioids without reducing analgesia.
- Chemogenetic stimulation of the hypoglossal neurons improves upper airway patency(2017) CURADO, Thomaz Fleury; FISHBEIN, Kenneth; Huy Pho; BRENNICK, Michael; DERGACHEVA, Olga; SENNES, Luiz U.; PHAM, Luu V.; LADENHEIM, Ellen E.; SPENCER, Richard; MENDELOWITZ, David; SCHWARTZ, Alan R.; POLOTSKY, Vsevolod Y.Obstructive sleep apnea (OSA) is characterized by recurrent upper airway obstruction during sleep. OSA leads to high cardiovascular morbidity and mortality. The pathogenesis of OSA has been linked to a defect in neuromuscular control of the pharynx. There is no effective pharmacotherapy for OSA. The objective of this study was to determine whether upper airway patency can be improved using chemogenetic approach by deploying designer receptors exclusively activated by designer drug (DREADD) in the hypoglossal motorneurons. DREADD (rAAV5-hSyn-hM3(Gq)-mCherry) and control virus (rAAV5-hSyn-EGFP) were stereotactically administered to the hypoglossal nucleus of C57BL/6J mice. In 6-8 weeks genioglossus EMG and dynamic MRI of the upper airway were performed before and after administration of the DREADD ligand clozapine-N-oxide (CNO) or vehicle (saline). In DREADD-treated mice, CNO activated the genioglossus muscle and markedly dilated the pharynx, whereas saline had no effect. Control virus treated mice showed no effect of CNO. Our results suggest that chemogenetic approach can be considered as a treatment option for OSA and other motorneuron disorders.
conferenceObject ACTIVATION OF LEPTIN RECEPTOR POSITIVE NEURONS IN THE NUCLEUS OF THE SOLITARY TRACT (NTS) ALLEVIATES SLEEP DISORDERED BREATHING IN OBESE MICE(2019) FREIRE, Carla; PHO, Huy; BERGER, Slava; FLEURY-CURADO, Thomaz; SENNES, Luiz U.; SCHWARTZ, Alan R.; POLOTSKY, Vsevolod Y.