key: cord-0961901-xsrkshoa authors: Naninck, Thibaut; Kahlaoui, Nidhal; Lemaitre, Julien; Maisonnasse, Pauline; De Mori, Antoine; Pascal, Quentin; Contreras, Vanessa; Marlin, Romain; Relouzat, Francis; Delache, Benoît; Hérate, Cécile; Aldon, Yoann; van Gils, Marit; Zabaleta, Nerea; Tsong Fang, Raphaël Ho; Bosquet, Nathalie; Sanders, Rogier W.; Vandenberghe, Luk H.; Chapon, Catherine; Le Grand, Roger title: Computed Tomography and [18F]-FDG PET imaging provide additional readouts for COVID-19 pathogenesis and therapies evaluation in non-human primates date: 2022-03-17 journal: iScience DOI: 10.1016/j.isci.2022.104101 sha: 5a1e8f47ca51f3c959e3158cb369983b548b5ede doc_id: 961901 cord_uid: xsrkshoa Non-human primates (NHPs) are particularly relevant as preclinical models for SARS-CoV-2 infection and nuclear imaging may represent a valuable tool for monitoring infection in this species. We investigated the benefit of computed X-ray tomography (CT) and [18F]-FDG positron-emission tomography (PET) to monitor the early phase of the disease in a large cohort (n=76) of SARS-CoV-2 infected macaques. Following infection, animals showed mild COVID-19 symptoms including typical lung lesions. CT scores at acute phase reflect the heterogeneity of lung burden following infection. Moreover, [18F]-FDG PET revealed that FDG uptake was significantly higher in the lungs, nasal cavities, lung-draining lymph nodes and spleen of NHPs by 5 days post infection compared to pre-infection levels, indicating early local inflammation. The comparison of CT and PET data from previous COVID-19 treatments or vaccine we tested in NHP, to this large cohort of untreated animals demonstrated the value of in vivo imaging in preclinical trials. The COVID-19 pandemic is still affecting millions of people worldwide and has been • Animal inclusion and disease monitoring 85 In total, 61 Mauritian cynomolgus macaques (CM) and 15 rhesus macaques (RM) were exposed 86 to the virus (or mock exposed as control) and included in our imaging longitudinal analysis, as 87 described in Table 1 Table S1 . Additional imaging data from our previous published studies 88 (Maisonnasse et al., 2021; Maisonnasse et al., 2020; Zabaleta et al., 2021) in CM assessing the 89 efficacy of diverse treatments (hydroxychloroquine, n = 5, monoclonal antibody, n = 5) or 90 vaccination (n = 6) against SARS-CoV-2 infection were then used to assess the strength of the 91 imaging readout (Table 2) . to 5.8 mm two days later), as already reported for humans (Albarello et al., 2020) . The same 115 lesion features could also be found in SARS-CoV-2-infected RM (Figure S1 E-F). 116 We performed semi-quantitative CT scoring to better characterize the lung-lesion burden, as Table S2 , reflects the lesion type and extension for each None of three mock-infected animals showed lung lesions (CT score = 0). The dose of viral 122 exposure did not appear to affect the CT scores. We observed no statistical difference during 123 J o u r n a l P r e -p r o o f the acute phase of infection (2-3 days post-infection (d.p.i.)) between animals exposed to 10 5 124 PFU and those exposed to 10 6 PFU (p = 0.40) for CM ( Figure 3A ). The two animals exposed 125 to 10 7 PFU also showed similar CT scores. Infected animals showed a broad range of lung 126 burden ( Figure 3A ). Most (n = 39/52, 75%) had mild lung lesions (CT scores < 5), but a few 127 showed larger lesions (extended GGOs and/or CPP). There was no statistical difference (p = 128 0.27) concerning the CT scores at 2-3 d.p.i. between cynomolgus and rhesus macaques exposed 129 to 10 5 PFU of SARS-CoV-2 ( Figure 3B ). Overall, NHP imaging CT data reflects the diversity 130 of human cases, with mostly mild to moderate cases (Oran and Topol, 2021) . The main objective was to assess hypermetabolism linked to inflammation and cell infiltration Figure S5B ). No major increase of 174 FDG uptake was detected in all these regions of interest in mock-infected animals ( Figure 6 ). Other organs of interest, such as the brain (1.09 ± 0.24) and liver (1.11 ± 0.24), did not show • Comparison of imaging data and virological or biological parameters 182 We first evaluated potential correlations between imaging data and viral RNA copy numbers in . We compared their CT scores to those of the 203 large cohort of control animals we described above (Table 2) . 204 There were no differences in the CT scores between HCQ-treated and control animals (p = 0.88, Limitations of the study 296 We provide imaging data on SARS-CoV-2 exposed animals with the initial "Wuhan" strain. CoV-2-exposed cynomolgus macaques according to the inoculum dose. Mock: exposed to PBS. (B) CT 349 scores at 2-3 d.p.i. in SARS-CoV-2 exposed (10 5 PFU) cynomolgus versus rhesus macaques. (C) 350 Evolution of the CT score between 2-3 d.p.i. and 10-14 d.p.i. for exposed cynomolgus macaques (*: Sixty-one cynomolgus macaques and 15 rhesus macaques, all young adults, were randomly 435 assigned between dose groups as described in Table S1 . All animals were then exposed to a 436 total dose of 10 5 , 10 6 , or 10 7 PFU of SARS-CoV-2 (BetaCoV/France/IDF/0372/2020, passaged 437 twice in VeroE6 cells) via a combination of the intranasal and the intratracheal routes (day 0), 438 using atropine (0.04 mg kg -1 ) for pre-medication and ketamine (5 mg kg -1 ) with medetomidine 439 (0.05 mg kg -1 ) for anaesthesia. Among them, three cynomolgus macaques were mock exposed 440 using saline with the same protocol. Animals were observed daily, and clinical exams were 441 performed at baseline, daily for one week, and then twice weekly on anaesthetized animals 442 using ketamine (5 mg kg -1 ) and medetomidine (0.05 mg kg -1 ). Nasopharyngeal specimens were and 3DSlicer (open-source tool). All CT lung images had the same window level of -300 and a window width of 1,600. Pulmonary lesions were defined as ground-glass opacity, crazy-paving pattern, or in the VOIs was given as a standardized uptake value (SUVmean, SUVmax) and organ-to-503 background ratios (or ratio to baseline) were calculated for each animal. 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