CARLA FREIRE DE CASTRO LIMA
Projetos de Pesquisa
Unidades Organizacionais
Instituto Central, Hospital das Clínicas, Faculdade de Medicina
LIM/63, Hospital das Clínicas, Faculdade de Medicina
LIM/63, Hospital das Clínicas, Faculdade de Medicina
6 resultados
Resultados de Busca
Agora exibindo 1 - 6 de 6
- 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.
bookPart Traqueostomia(2019) LIMA, Carla Freire de Castro- 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.
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.- Treatment of Sleep Disordered Breathing with Leptin Loaded Extracellular Vesicles(2022) FREIRE, Carla; PHO, Huv; RAMSEV, Jacob D.; ZHAO, Yuling; KIM, Lenise J.; BERGER, Slava; ANOKYE-DANSO, Frederick; SENNES, Luiz U.; AHIMA, Rexford S.; BATRAKOVA, Elena V.; KABANOV, Alexander V.; POLOTSKY, Vsevolod Y.