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Background: Posterior sagittal is a very well accepted approach in the treatment of anorectal malformations. This approach provides good access and exposure through the perineum to the deep pelvic structures. It reduces risk of injury to important structures as dissection remains in midline. Aims and Objectives: To access feasibility of posterior sagittal approach for non-anorectal malformation indications and to widen the spectrum. Material and Methods: We present a series of 10 cases of non-anorectal malformations operated by this approach for 4 years. Results: Six patients included in the study were of Disorders of Sexual Differentiation with pseudovagina, three of Y duplication of the urethra, and one of cervical atresia. All patients had good results. Conclusion: Posterior sagittal approach is feasible, safe with minimal bleeding, and no postoperative incontinence. It can safely be used for non-anorectal indications.
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Introduction: Hernia of the umbilical cord (HUC) is an uncommon problem which is often misdiagnosed, leading to inadequate treatment and various complications which cause increased morbidity and mortality in neonates. To address this issue, we took up this study. Materials and Methods: Patients diagnosed with HUC from January 1, 2017, to December 31, 2021, were retrospectively analyzed. The following data of all these patients were collected and retrospectively analyzed: demography, radiological investigations, echocardiography, contents of hernia, type of surgery performed, and outcome. Results: Eighteen out of 19 patients included in the study were taken up for surgery after baseline investigations and echocardiography. Out of 19 patients, 15 were discharged successfully and are on regular follow-up. Of the remaining four patients, an anastomotic leak occurred in two and they went into sepsis and succumbed. One of the patients died before any intervention and one succumbed on 3rd postoperative day due to sepsis. Conclusion: Timely referral and intervention can save precious lives. We need to educate doctors and health-care providers so that proper diagnosis and timely management can be done for this anomaly which is associated with less morbidity and a lower rate of associated anomalies.
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Introduction: Peutz-Jeghers Syndrome (PJS) is an autosomal dominant disease presenting with hamartomatous polyps in the gastrointestinal tract and hyperpigmented macules on lips and oral mucosa. The incidence of this syndrome is approximately 1 in 1,20,000 births. Materials and Methods: In this article, we are presenting 11 cases of PJS which were misdiagnosed and patients were compelled to visit hospital repeatedly. All these cases were diagnosed based on clinical suspicion, family history, and histopathological examination of specimens. Most of the cases presented with intussusception and required emergency surgical management. Results: PJS can be diagnosed by the presence of microscopically confirmed hamartomatous polyps and a minimum of two of the following clinical criteria: Family history, mucocutaneous melanotic spots, and small bowel polyps with bleeding per rectally. The diagnosis can be missed if the melanotic spots on the face are missed. Routine investigations, imaging, and endoscopy were done in all cases. PJS patients need regular follow-up due to chance of recurrence of symptoms and susceptibility to cancer. Conclusion: PJS needs a high index of suspicion for diagnosis in cases of recurrent abdominal pain with bleeding per rectum. Proper family history and meticulous clinical examination for melanosis are very important to prevent the misdiagnosis of these cases.
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We report an infant reared as a girl who presented with ambiguous genitalia and urine coming per rectum. On examination, she had minimal clitoromegaly with labial fusion and Y urethral duplication. On investigations, genetic, metabolic, and endocrine causes for disorder of sexual differentiation were ruled out. This girl was operated via a posterior sagittal approach. This case was unique in that the urogenital sinus was deviated posteriorly and opened in the anterior rectal wall and the accessory urethra opened in clitoris, along with ambiguity of external genitalia, thus making it a rare variant of the posterior cloaca. The diagnosis requires a high index of suspicion in clinician and a meticulous examination of the external genitalia. Very little literature is available for this rare anomaly. Few reports have classified them as female pseudohermaphroditism with cloacal and urogenital sinus defects. Hence, we hereby discuss and review the literature for previously reported cases.
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Therapeutic strategies based on in vitro-transcribed mRNA (IVT) are attractive because they avoid the permanent signature of genomic integration that is associated with DNA-based therapy and result in the transient production of proteins of interest. To date, IVT has mainly been used in vaccination protocols to generate immune responses to foreign Ags. In this "proof-of-principle" study, we explore a strategy of combinatorial IVT to recruit and reprogram immune effector cells to acquire divergent biological functions in mice in vivo. First, we demonstrate that synthetic mRNA encoding CCL3 is able to recruit murine monocytes in a nonprogrammed state, exhibiting neither bactericidal nor tissue-repairing properties. However, upon addition of either Ifn-γ mRNA or Il-4 mRNA, we successfully polarized these cells to adopt either M1 or M2 macrophage activation phenotypes. This cellular reprogramming was demonstrated through increased expression of known surface markers and through the differential modulation of NADPH oxidase activity, or the superoxide burst. Our study demonstrates how IVT strategies can be combined to recruit and reprogram immune effector cells that have the capacity to fulfill complex biological tasks in vivo.
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Reprogramação Celular , Macrófagos/imunologia , Monócitos/imunologia , RNA Mensageiro/imunologia , Animais , Células Cultivadas , Quimiocina CCL2/genética , Quimiocina CCL2/imunologia , Quimiocina CCL3/genética , Células HeLa , Humanos , Interferon gama/genética , Interleucina-4/genética , Linfócitos/imunologia , Ativação de Macrófagos , Camundongos , Camundongos Endogâmicos C57BL , Estudo de Prova de Conceito , RNA Mensageiro/síntese química , Transcrição GênicaRESUMO
There is a clear need for low-cost, self-applied, long-lasting approaches to prevent human immunodeficiency virus (HIV) infection in both men and women, even with the advent of pre-exposure prophylaxis (PrEP). Broadly neutralizing antibodies represent an option to improve HIV prophylaxis, but intravenous delivery, cold-chain stability requirements, low cervicovaginal concentrations, and cost may preclude their use. Here, we present an approach to express the anti-GP120 broadly neutralizing antibody PGT121 in the primary site of inoculation, the female reproductive tract, using synthetic mRNA. Expression is achieved through aerosol delivery of unformulated mRNA in water. We demonstrated high levels of antibody expression for over 28 days with a single mRNA administration in the reproductive tract of sheep. In rhesus macaques, neutralizing antibody titers in secretions developed within 4 h and simian-HIV (SHIV) infection of ex vivo explants was prevented. Persistence of PGT121 in vaginal secretions and epithelium was achieved through the incorporation of a glycosylphosphatidylinositol (GPI) anchor into the heavy chain of the antibody. Overall, we present a new paradigm to deliver neutralizing antibodies to the female reproductive tract for the prevention of HIV infections.
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Anticorpos Amplamente Neutralizantes/imunologia , Expressão Gênica , Anticorpos Anti-HIV/imunologia , Mucosa/imunologia , Mucosa/metabolismo , RNA Mensageiro/administração & dosagem , Vagina , Vacinas contra a AIDS/administração & dosagem , Vacinas contra a AIDS/genética , Vacinas contra a AIDS/imunologia , Aerossóis , Animais , Chlorocebus aethiops , Feminino , Imunofluorescência , Infecções por HIV/imunologia , HIV-1/imunologia , Camundongos , Testes de Neutralização , RNA Mensageiro/síntese química , Ovinos , Síndrome de Imunodeficiência Adquirida dos Símios/imunologia , Vírus da Imunodeficiência Símia/imunologia , Vagina/imunologia , Vagina/metabolismo , Células VeroRESUMO
Dysfunctional endothelium causes more disease than any other cell type. Systemically administered RNA delivery to nonliver tissues remains challenging, in large part because there is no high-throughput method to identify nanoparticles that deliver functional mRNA to cells in vivo. Here we report a system capable of simultaneously quantifying how >100 lipid nanoparticles (LNPs) deliver mRNA that is translated into functional protein. Using this system (named FIND), we measured how >250 LNPs delivered mRNA to multiple cell types in vivo and identified 7C2 and 7C3, two LNPs that efficiently deliver siRNA, single-guide RNA (sgRNA), and mRNA to endothelial cells. The 7C3 delivered Cas9 mRNA and sgRNA to splenic endothelial cells as efficiently as hepatocytes, distinguishing it from LNPs that deliver Cas9 mRNA and sgRNA to hepatocytes more than other cell types. These data demonstrate that FIND can identify nanoparticles with novel tropisms in vivo.
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Sistemas CRISPR-Cas , Células Endoteliais/metabolismo , Edição de Genes , Técnicas de Transferência de Genes , Lipídeos/química , Nanopartículas/administração & dosagem , RNA Guia de Cinetoplastídeos/genética , RNA Mensageiro/genética , Animais , Células Cultivadas , Células Endoteliais/citologia , Células HEK293 , Hepatócitos/citologia , Hepatócitos/metabolismo , Ensaios de Triagem em Larga Escala , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Nanopartículas/química , RNA Guia de Cinetoplastídeos/química , RNA Mensageiro/químicaRESUMO
BACKGROUND: Appropriately modulating inflammation after traumatic brain injury (TBI) may prevent disabilities for the millions of those inflicted annually. In TBI, cellular mediators of inflammation, including macrophages and microglia, possess a range of phenotypes relevant for an immunomodulatory therapeutic approach. It is thought that early phenotypic modulation of these cells will have a cascading healing effect. In fact, an anti-inflammatory, "M2-like" macrophage phenotype after TBI has been associated with neurogenesis, axonal regeneration, and improved white matter integrity (WMI). There already exist clinical trials seeking an M2-like bias through mesenchymal stem/stromal cells (MSCs). However, MSCs do not endogenously synthesize key signals that induce robust M2-like phenotypes such as interleukin-4 (IL-4). METHODS: To enrich M2-like macrophages in a clinically relevant manner, we augmented MSCs with synthetic IL-4 mRNA to transiently express IL-4. These IL-4 expressing MSCs (IL-4 MSCs) were characterized for expression and functionality and then delivered in a modified mouse TBI model of closed head injury. Groups were assessed for functional deficits and MR imaging. Brain tissue was analyzed through flow cytometry, multi-plex ELISA, qPCR, histology, and RNA sequencing. RESULTS: We observed that IL-4 MSCs indeed induce a robust M2-like macrophage phenotype and promote anti-inflammatory gene expression after TBI. However, here we demonstrate that acute enrichment of M2-like macrophages did not translate to improved functional or histological outcomes, or improvements in WMI on MR imaging. To further understand whether dysfunctional pathways underlie the lack of therapeutic effect, we report transcriptomic analysis of injured and treated brains. Through this, we discovered that inflammation persists despite acute enrichment of M2-like macrophages in the brain. CONCLUSION: The results demonstrate that MSCs can be engineered to induce a stronger M2-like macrophage response in vivo. However, they also suggest that acute enrichment of only M2-like macrophages after diffuse TBI cannot orchestrate neurogenesis, axonal regeneration, or improve WMI. Here, we also discuss our modified TBI model and methods to assess severity, behavioral studies, and propose that IL-4 expressing MSCs may also have relevance in other cavitary diseases or in improving biomaterial integration into tissues.
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Lesões Encefálicas Traumáticas/metabolismo , Interleucina-4/metabolismo , Macrófagos/metabolismo , Células-Tronco Mesenquimais/metabolismo , Animais , Modelos Animais de Doenças , Inflamação/metabolismo , Masculino , Camundongos , Microglia/metabolismoRESUMO
Cervical atresia is a rare association with anorectal malformation (ARM) which can be missed till puberty in the presence of normal vaginal orifice. A 12-year-old girl operated for ARM in neonatal age presented with primary amenorrhea. She had a normal vaginal opening, short perineal body, and prolapsed anteposed anus and was diagnosed with cervical agenesis. As the posterior sagittal approach is standard to place the rectum in correct anatomical position, reconstruction of the anus along with adequate perineal body and uterovaginal anastomosis was performed through this approach. This report highlights the utility and versatility of this approach for the management of such complex cases.
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BACKGROUND/PURPOSE: The aim of our study is to determine efficacy, safety, and feasibility of video-assisted thoracoscopic surgery (VATS) in childhood empyema with a technique of only two ports and open instruments at a tertiary care center in India. METHODS: This is a retrospective study of patients below 18 years, with empyema presenting to the Department of Pediatric Surgery of a Tertiary Care Referral Hospital in India, over a period of 9 years who underwent VATS decortication. Only two ports with open surgical instruments were used. The patients were assessed on the basis of mean duration of preoperative symptoms, duration of surgery, average blood loss, postoperative pain relief, complications, and need for redo surgery. RESULTS: A total of 97 patients underwent primary VATS decortications without inserting an intercostal drainage (ICD) tube and 70 patients as a secondary procedure after ICD tube was inserted. Mean duration of symptoms was 11 days. The average blood loss during surgery was estimated to be 170 cc. The mean duration of surgery was 90 min. The most common postoperative complication was air leak seen in 19.16% of patients. Minor leaks usually settled by 24 h. In eight patients, a negative suction had to be applied to the ICD tube for persistent air leak. The average length of postoperative stay was 4 days. Two patients required a repeat open decortication procedure due to nonresolution of symptoms and poor lung expansion after VATS. Patients had minimal pain and excellent cosmetic outcome after VATS. CONCLUSION: Two-port VATS decortication procedure is as feasible and effective as three-port procedure for decortication with better cosmetic result and pain relief.
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Respiratory syncytial virus (RSV) causes severe pneumonia and bronchiolitis in infants, children and older adults. The use of metallic nanoparticles as potential therapeutics is being explored against respiratory viruses like Influenza, Parainfluenza and Adenovirus. In this study, we showed that gold nanorods (GNRs) inhibit RSV in HEp-2 cells and BALB/c mice by 82% and 56%, respectively. The RSV inhibition correlated with marked upregulated antiviral genes due to GNR mediated TLR, NOD-like receptor and RIG-I-like receptor signaling pathways. Transmission electron microscopy of lungs showed GNRs in the endocytotic vesicles and histological analyses indicated infiltration by neutrophils, eosinophils and monocytes correlating with clearance of RSV. In addition, production of cytokines and chemokines in the lungs indicates recruitment of immune cells to counter RSV replication. To our knowledge, this is the first in vitro and in vivo report that provides possible antiviral mechanisms of GNRs against RSV.
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Ouro/farmacologia , Imunidade Inata , Nanotubos , Infecções por Vírus Respiratório Sincicial/tratamento farmacológico , Vírus Sinciciais Respiratórios/efeitos dos fármacos , Animais , Ouro/administração & dosagem , Humanos , Camundongos , Camundongos Endogâmicos NODRESUMO
Tuberculosis is the most frequent cause of death, specifically caused by a single infectious agent, Mycobacterium tuberculosis. There are two types of tuberculosis: pulmonary tuberculosis and extrapulmonary tuberculosis. Patients with extrapulmonary tuberculosis often have reduced lung function due to the disease's structural abnormalities, which also significantly impair their quality of life. The suggested standard of care for the treatment of extrapulmonary tuberculosis patients is pulmonary rehabilitation. A 35-year-old male patient who complained of shortness of breath, dry cough, and on-and-off fever diagnose with extrapulmonary tuberculosis was the subject of the case study. The patient had extrapulmonary tuberculosis with a history of pleural effusion, which was managed with proper medications. After increasing symptoms of the disease, the patient was referred for pulmonary rehabilitation. Physiotherapy protocol includes breathing exercises, relaxation techniques, and mobility exercises for the upper limb and lower limb. Effective physical rehabilitation was necessary to minimize complications and allow him to resume daily activities. Several outcome measures, like the dyspnea scale, visual analog scale, six-minute walk test, and World Health Organization-Quality of Life (WHO-QOL) questionnaire, were used to monitor the patient's progress during rehabilitation. The benefits of physiotherapy protocols emphasize the need for tailored approaches to addressing individual patient needs for comprehensive recovery as it significantly enhances clinical, physical, psychosocial, and overall quality of life, making it crucial for patients with extrapulmonary tuberculosis. The protocols are beneficial to improve exercise capacity, muscle force, symptoms such as dyspnea, cough, and health-related quality of life in these patients. In this study, the focus was more on breathing exercises such as segmental breathing exercises for lung expansion and increasing air entry in the lungs followed by improving functional capacity and strength.
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Introduction: The presentation of macroprolactinomas and response to treatment may vary according to age, sex and tumour characteristics. To analyse clinical phenotype, biochemical and radiological characteristics of macroprolactinomas presenting to a tertiary care centre. A retrospective observational study from January 2018 to December 2022. Methods: Thirty diagnosed cases (18 females, 12 males) of macroprolactinomas were included and followed up for one year. Results: The most common presentation was headache (73%), visual disturbances (50%), galactorrhoea (33.3%) and loss of libido (26.6%) along with menstrual cycle disturbances (94%), and infertility (55%) in females. Duration of symptoms (2.22 ± 2.87 vs 4.61 ± 3.4 years), tumour size (4.8 ± 2.09 vs 2.75 ± 1.24 cm) and prolactin levels (5153.5 ± 4755.3 vs 1803.5 ± 3785.5 ng/ml) were different significantly between males and females. Good response to medical therapy was observed in 84% of the treatment-naive patients. Conclusion: Macroprolactinomas in males present with shorter duration of symptoms, larger size, higher prolactin levels and more resistant tumours, emphasizing the need for early diagnosis and aggressive management. Medical therapy remains the treatment of choice irrespective of gender.
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Osteoarthritis (OA) of the hip is a rare condition that occurs in adults can be a result of avascular necrosis or a history of steroids that can wear away the articulating cartilage of the hip joint causing friction, pain in the groin region, stiffness, and decreased functional mobility. We present a 30-year-old adult who came with chief complaints of pain in the groin region, stiffness, difficulty in walking, and experiencing pain while walking, which had reduced his activity of daily living. The investigation was done, and the patient was diagnosed with bilateral hip OA secondary to avascular necrosis. To reduce morbidity, preoperative physiotherapy management for eight weeks was planned and started before the operation. The purpose was to educate the patient about the condition, reduce pain, increase the ranges of the hip, improve strength, and provide gait re-education. We added basic proprioception training and plyometric exercises for the hip to improve strength and balance. At the end of the session patient, positive results were achieved. The progress of proprioception or balance training can be improved by using single-leg balance as an outcome measure. Hence, our study aims to use exercise therapy to reduce or postpone the need for hip arthroscopy. However, future research should focus on plyometric exercises for the lower limbs or any abnormalities associated with the lower limbs. However, they should be carried out when some recovery is observed in patients.
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Syringomyelia is a center-medullary syndrome characterized by the presence of fluid-filled spaces known as syrinx within the spinal canal. Arnold Chiari Malformation (CM-I), a rhombencephalon anomaly formerly identified as hindbrain hernia, is usually associated with it. This disorder causes the brain (cerebellum) to bulge through the opening in the skull known as the foramen magnum. Some asymptomatic patients may develop symptoms quickly if they jolt their heads and cough for a lengthy period of time. Syringomyelia can be caused by trauma, illness, inflammation, or previous surgery that affects the circulation of cerebral spinal fluid resulting in CSF flow obstruction. The discomfort is acute and progressive, radiating to the neck and shoulder, and is accompanied by sensory loss, motor atrophy, decreased hearing, oscillopsia, and cerebellar abnormalities. This case report is of a 39-year-old woman diagnosed with syringomyelia associated with Arnold Chiari malformation and showed similar symptoms managed by foramen decompression and tonsillar elevation surgery. It involves removing a small piece of bone from the skull and a small section of the 1st vertebra from the back of the neck and head. In this way, there is an increase in skull space. Decompression of the spinal canal increases the size of the subarachnoid cisterns and constricts the syrinx cavity. After surgery, physiotherapy was advised because all superficial sensations over C8 and T1 were diminished, the range of motion along with strength was reduced, doing daily activities was difficult, and quality of life was affected. So, by decreasing symptoms and improving the patient's quality of life, physiotherapy improved the patient's condition significantly in this case report. The rationale of this study is to show the importance of physiotherapy in recovering after a neurological condition followed by corrective neurosurgery.
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Primary patellar dislocation or first-time patellar dislocation is the second most frequent cause of knee injuries which overall accounts for about 3% of other knee injuries. The patellofemoral joint is formed by the patella connecting to the femoral trochlea and creates both static and dynamic structures of the knee. There are basically three types of patellar dislocation: superior, lateral, and medial. The lateral dislocation is the most frequent one. Females are more vulnerable and are at higher risk than males. Muscular weakness or muscular imbalance leads to patellar instability, and ultimately to dislocation. The recurrence rate after primary patellar dislocation is 15-60%. This case report is of a 31-year-old female with patella dislocation with a medial meniscal tear and a case of early osteoarthritis for whom we planned goal-oriented physiotherapy rehabilitation week-wise and progressed every week. The assessment was taken before and after physiotherapy rehabilitation. The patient was managed conservatively with a long knee brace, and physiotherapy started after one month. Due to prolonged immobilization, the patient suffered from quadriceps muscle atrophy. The physiotherapist focused on biomechanism and got the expected results in pain reduction, regaining strength, and improving range of motion, and the patient was able to walk properly without taking any support after rehabilitation.
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This paper describes the simulation by Solar Cell Capacitance Simulator-1D (SCAPS-1D) software of ZnO/CdS/SnS/NiO/Au solar cells, in which zinc oxide (ZnO) is used as transparent conductive oxide (TCO) and nickel oxide (NiO) is used as a hole transport layer (HTL). The effects of absorber layer (SnS) thickness, carrier concentration, SnS defect density, NiO HTL, ZnO TCO, electron affinity and work function on cell performance have been evaluated. The effect of interface defect density of SnS/CdS on the performance of the heterojunction solar cell is also analysed. As the results indicate, a maximum power conversion efficiency of 26.92% was obtained.
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In the present paper, the theoretical investigation of the device structure ITO/CeO2/SnS/Spiro-OMeTAD/Mo of SnS-based solar cell has been performed. The aim of this work is to examine how the Spiro-OMeTAD HTL affects the performance of SnS-based heterostructure solar cell. Using SCAPS-1D simulation software, various parameters of SnS-based solar cell such as work function, series and shunt resistance and working temperature have been investigated. With the help of Spiro-OMeTAD, the suggested cell's open-circuit voltage was increased to 344 mV. The use of Spiro-OMeTAD HTL in the SnS-based solar cell resulted in 14% efficiency increase, and the proposed heterojunction solar cell has 25.65% efficiency. The cell's performance is determined by the carrier density and width of the CeO2 ETL (electron transport layer), SnS absorber layer and Spiro-OMeTAD HTL (hole transport layer). These data reveal that the Spiro-OMeTAD solar cells could have been a good HTL (hole transport layer) in regards to producing SnS-based heterojunction solar cell with high efficiency and reduced cost.
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The search for a preventive vaccine against HIV infection remains an ongoing challenge, indicating the need for novel approaches. Parainfluenza virus 5 (PIV5) is a paramyxovirus replicating in the upper airways that is not associated with any animal or human pathology. In animal models, PIV5-vectored vaccines have shown protection against influenza, RSV, and other human pathogens. Here, we generated PIV5 vaccines expressing HIV envelope (Env) and SIV Gag and administered them intranasally to macaques, followed by boosting with virus-like particles (VLPs) containing trimeric HIV Env. Moreover, we compared the immune responses generated by PIV5-SHIV prime/VLPs boost regimen in naïve vs a control group in which pre-existing immunity to the PIV5 vector was established. We demonstrate for the first time that intranasal administration of PIV5-based HIV vaccines is safe, well-tolerated and immunogenic, and that boosting with adjuvanted trimeric Env VLPs enhances humoral and cellular immune responses. The PIV5 prime/VLPs boost regimen induced robust and durable systemic and mucosal Env-specific antibody titers with functional activities including ADCC and neutralization. This regimen also induced highly polyfunctional antigen-specific T cell responses. Importantly, we show that diminished responses due to PIV5 pre-existing immunity can be overcome in part with VLP protein boosts. Overall, these results establish that PIV5-based HIV vaccine candidates are promising and warrant further investigation including moving on to primate challenge studies.
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Vacinas contra a AIDS/administração & dosagem , Produtos do Gene gag/administração & dosagem , HIV-1/imunologia , Imunogenicidade da Vacina , Vírus da Parainfluenza 5/imunologia , Vírus da Imunodeficiência Símia/imunologia , Vírion/imunologia , Produtos do Gene env do Vírus da Imunodeficiência Humana/administração & dosagem , Vacinas contra a AIDS/genética , Vacinas contra a AIDS/imunologia , Administração Intranasal , Animais , Anticorpos Antivirais/sangue , Bovinos , Linhagem Celular , Produtos do Gene gag/genética , Produtos do Gene gag/imunologia , HIV-1/genética , Interações Hospedeiro-Patógeno , Imunidade Celular , Imunidade Humoral , Imunidade nas Mucosas , Macaca mulatta , Masculino , Mucosa Nasal/imunologia , Mucosa Nasal/virologia , Vírus da Parainfluenza 5/genética , Vírus da Imunodeficiência Símia/genética , Linfócitos T/imunologia , Linfócitos T/virologia , Vacinação , Vacinas de DNA/administração & dosagem , Vacinas de DNA/imunologia , Vírion/genética , Produtos do Gene env do Vírus da Imunodeficiência Humana/genética , Produtos do Gene env do Vírus da Imunodeficiência Humana/imunologiaRESUMO
Cas13a has been used to target RNA viruses in cell culture, but efficacy has not been demonstrated in animal models. In this study, we used messenger RNA (mRNA)-encoded Cas13a for mitigating influenza virus A and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in mice and hamsters, respectively. We designed CRISPR RNAs (crRNAs) specific for PB1 and highly conserved regions of PB2 of influenza virus, and against the replicase and nucleocapsid genes of SARS-CoV-2, and selected the crRNAs that reduced viral RNA levels most efficiently in cell culture. We delivered polymer-formulated Cas13a mRNA and the validated guides to the respiratory tract using a nebulizer. In mice, Cas13a degraded influenza RNA in lung tissue efficiently when delivered after infection, whereas in hamsters, Cas13a delivery reduced SARS-CoV-2 replication and reduced symptoms. Our findings suggest that Cas13a-mediated targeting of pathogenic viruses can mitigate respiratory infections.