{ "items": [ "\n\n
<p>Supplementary Materials and Methods, Supplementary Figures S1-S12</p>
\n \n\n \n \n<div>Abstract<p>A lack of models that recapitulate the complexity of human bone marrow has hampered mechanistic studies of normal and malignant hematopoiesis and the validation of novel therapies. Here, we describe a step-wise, directed-differentiation protocol in which organoids are generated from induced pluripotent stem cells committed to mesenchymal, endothelial, and hematopoietic lineages. These 3D structures capture key features of human bone marrow\u2014stroma, lumen-forming sinusoids, and myeloid cells including proplatelet-forming megakaryocytes. The organoids supported the engraftment and survival of cells from patients with blood malignancies, including cancer types notoriously difficult to maintain <i>ex vivo</i>. Fibrosis of the organoid occurred following TGF\u03b2 stimulation and engraftment with myelofibrosis but not healthy donor\u2013derived cells, validating this platform as a powerful tool for studies of malignant cells and their interactions within a human bone marrow\u2013like milieu. This enabling technology is likely to accelerate the discovery and prioritization of novel targets for bone marrow disorders and blood cancers.</p>Significance:<p>We present a human bone marrow organoid that supports the growth of primary cells from patients with myeloid and lymphoid blood cancers. This model allows for mechanistic studies of blood cancers in the context of their microenvironment and provides a much-needed <i>ex vivo</i> tool for the prioritization of new therapeutics.</p><p><i><a href=\"https://aacrjournals.org/cancerdiscovery/article/doi/10.1158/2159-8290.CD-22-1303\" target=\"_blank\">See related commentary by Derecka and Crispino, p. 263</a>.</i></p><p><i><a href=\"https://aacrjournals.org/cancerdiscovery/article/doi/10.1158/2159-8290.CD-13-2-ITI\" target=\"_blank\">This article is highlighted in the In This Issue feature, p. 247</a></i></p></div>
\n \n\n \n \n<p>Supplementary Table 2 Gene sets for GSEA</p>
\n \n\n \n \n<p>Supplementary Table 5 Antibodies</p>
\n \n\n \n \n<p>Supplementary Table 2 Gene sets for GSEA</p>
\n \n\n \n \n<p>Supplementary Table 1 VEGFAC Top Differentially Expressed Genes by Cluster</p>
\n \n\n \n \n<p>MM ALL Donor Details</p>
\n \n\n \n \n<p>Supplementary Table 6 qRT PCR Probes</p>
\n \n\n \n \n<p>Supplementary Table 5 Antibodies</p>
\n \n\n \n \n<p>Supplementary Table 7 NGS Panel</p>
\n \n\n \n \n<p>Supplementary Table 3 HD and MPN samples.</p>
\n \n\n \n \n<p>MM ALL Donor Details</p>
\n \n\n \n \n<p>Supplementary Table 3 HD and MPN samples.</p>
\n \n\n \n \n<p>Supplementary Materials and Methods, Supplementary Figures S1-S12</p>
\n \n\n \n \n<p>Supplementary Table 7 NGS Panel</p>
\n \n\n \n \n<div>Abstract<p>A lack of models that recapitulate the complexity of human bone marrow has hampered mechanistic studies of normal and malignant hematopoiesis and the validation of novel therapies. Here, we describe a step-wise, directed-differentiation protocol in which organoids are generated from induced pluripotent stem cells committed to mesenchymal, endothelial, and hematopoietic lineages. These 3D structures capture key features of human bone marrow\u2014stroma, lumen-forming sinusoids, and myeloid cells including proplatelet-forming megakaryocytes. The organoids supported the engraftment and survival of cells from patients with blood malignancies, including cancer types notoriously difficult to maintain <i>ex vivo</i>. Fibrosis of the organoid occurred following TGF\u03b2 stimulation and engraftment with myelofibrosis but not healthy donor\u2013derived cells, validating this platform as a powerful tool for studies of malignant cells and their interactions within a human bone marrow\u2013like milieu. This enabling technology is likely to accelerate the discovery and prioritization of novel targets for bone marrow disorders and blood cancers.</p>Significance:<p>We present a human bone marrow organoid that supports the growth of primary cells from patients with myeloid and lymphoid blood cancers. This model allows for mechanistic studies of blood cancers in the context of their microenvironment and provides a much-needed <i>ex vivo</i> tool for the prioritization of new therapeutics.</p><p><i><a href=\"https://aacrjournals.org/cancerdiscovery/article/doi/10.1158/2159-8290.CD-22-1303\" target=\"_blank\">See related commentary by Derecka and Crispino, p. 263</a>.</i></p><p><i><a href=\"https://aacrjournals.org/cancerdiscovery/article/doi/10.1158/2159-8290.CD-13-2-ITI\" target=\"_blank\">This article is highlighted in the In This Issue feature, p. 247</a></i></p></div>
\n \n\n \n \nBACKGROUND: BCG confers reduced, variable protection against pulmonary tuberculosis. A more effective vaccine is needed. We evaluated the safety and immunogenicity of candidate regimen ChAdOx1 85A-MVA85A compared with BCG revaccination among Ugandan adolescents. METHODS: After ChAdOx1 85A dose escalation and age de-escalation, we did a randomised open-label phase 2a trial among healthy adolescents aged 12-17 years, who were BCG vaccinated at birth, without evident tuberculosis exposure, in Entebbe, Uganda. Participants were randomly assigned (1:1) using a block size of 6, to ChAdOx1 85A followed by MVA85A (on day 56) or BCG (Moscow strain). Laboratory staff were masked to group assignment. Primary outcomes were solicited and unsolicited adverse events (AEs) up to day 28 and serious adverse events (SAEs) throughout the trial; and IFN-\u03b3 ELISpot response to antigen 85A (day 63 [geometric mean] and days 0-224 [area under the curve; AUC). FINDINGS: Six adults (group 1, n=3; group 2, n=3) and six adolescents (group 3, n=3; group 4, n=3) were enrolled in the ChAdOx1 85A-only dose-escalation and age de-escalation studies (July to August, 2019). In the phase 2a trial, 60 adolescents were randomly assigned to ChAdOx1 85A-MVA85A (group 5, n=30) or BCG (group 6, n=30; December, 2019, to October, 2020). All 60 participants from groups 5 and 6 were included in the safety analysis, with 28 of 30 from group 5 (ChAdOx1 85A-MVA85A) and 29 of 30 from group 6 (BCG revaccination) analysed for immunogenicity outcomes. In the randomised trial, 60 AEs were reported among 23 (77%) of 30 participants following ChAdOx1 85A-MVA85A, 31 were systemic, with one severe event that occurred after the MVA85A boost that was rapidly self-limiting. All 30 participants in the BCG revaccination group reported at least one mild to moderate solicited AE; most were local reactions. There were no SAEs in either group. Ag85A-specific IFN-\u03b3 ELISpot responses peaked on day 63 in the ChAdOx1 85A-MVA85A group and were higher in the ChAdOx1 85A-MVA85A group compared with the BCG revaccination group (geometric mean ratio 30\u00b759 [95% CI 17\u00b746-53\u00b759], p<0\u00b70001, day 63; AUC mean difference 57\u2009091 [95% CI 40\u2009524-73\u2009658], p<0\u00b70001, days 0-224). INTERPRETATION: The ChAdOx1 85A-MVA85A regimen was safe and induced stronger Ag85A-specific responses than BCG revaccination. Our findings support further development of booster tuberculosis vaccines. FUNDING: UK Research and Innovations and Medical Research Council. TRANSLATIONS: For the Swahili and Luganda translations of the abstract see Supplementary Materials section.
\n \n\n \n \nThe Covid-19 pandemic is a unique event in the modern history of humanity, which has generated great challenges and, at the same time, valuable opportunities for public health. A number of examples of them are found in the more than three hundred pages of this book. In each chapter it is possible to understand how a vaccine for Covid-19 was developed in record time - due to the urgency of an antidote that would allow us to deal with this terrible disease - through the acceleration, compliance and improvement of all labor criteria, production, evaluation, timely release, and security. This entire process of developing the first vaccine produced by Brazil was described in a very creative way, allowing the reader to dive into a technical-scientific content of the highest level. The book presents an overview that goes through the origin of the virus, the transmission mechanisms of SARS-CoV-2, the vaccine development process and the regulatory and legal instruments to guarantee access to vaccination - starting with the most vulnerable populations. It also describes the trials and phases of clinical study development that ensured the vaccine's safety and efficacy. It also covers the logistics of distribution and pharmacovigilance for monitoring the product in the user population until the detailing of the technological prospection, as well as showing the necessary steps to carry out a process of technology transfer of the vaccine from the viral vector. Among the various innovations, it is worth highlighting: preparation of a technological order through ETEC; use of continuous submission to the National Health Surveillance Agency (ANVISA); and emergency use authorization. This effort made it possible to meet the expressive demand for Covid-19 vaccines in Brazil in a timely manner and on an unprecedented scale. For the Pan American Health Organization (PAHO), which turns 120 on December 2, 2022, it is an honor to have Bio-Manguinhos/Fiocruz as part of our history. Due to its outstanding performance on the international scene, this institute is a true heritage of humanity. And now, with the first Brazilian vaccine for Covid-19, Bio-Manguinhos/Fiocruz consolidates Brazil's leadership in the production of immunobiologicals in Latin America and the Caribbean, ensuring greater self-sufficiency and sustainability of basic health supplies not only for the country, but for the entire region of the Americas.
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