Uma capitania dos novos tempos: economia, sociedade e política na São Paulo restaurada (1765-1822)

O artigo reflete sobre a trajetória da Capitania de São Paulo, a partir de 1750, apontando sua transformação, de fronteira e "boca do sertão", para território estratégico da conquista e defesa das partes meridionais e área economicamente integrada aos circuitos mercantis atlânticos.In this article, we reflect upon the history of the Captaincy of São Paulo as from 1750, drawing attention to its transformation from frontier land and "door to the backcountry" into a territory of strategic value for the purposes of conquest and defense of the southern regions, and economically integrated into the Atlantic trade routes

Engineering NK-CAR.19 cells with the IL-15/IL-15Rα complex improved proliferation and anti-tumor effect in vivo

IntroductionNatural killer 92 (NK-92) cells are an attractive therapeutic approach as alternative chimeric antigen receptor (CAR) carriers, different from T cells, once they can be used in the allogeneic setting. The modest in vivo outcomes observed with NK-92 cells continue to present hurdles in successfully translating NK-92 cell therapies into clinical applications. Adoptive transfer of CAR-NK-92 cells holds out the promise of therapeutic benefit at a lower rate of adverse events due to the absence of GvHD and cytokine release syndrome. However, it has not achieved breakthrough clinical results yet, and further improvement of CAR-NK-92 cells is necessary.MethodsIn this study, we conducted a comparative analysis between CD19-targeted CAR (CAR.19) co-expressing IL-15 (CAR.19-IL15) with IL-15/IL-15Rα (CAR.19-IL15/IL15Rα) to promote NK cell proliferation, activation, and cytotoxic activity against B-cell leukemia. CAR constructs were cloned into lentiviral vector and transduced into NK-92 cell line. Potency of CAR-NK cells were assessed against CD19-expressing cell lines NALM-6 or Raji in vitro and in vivo in a murine model. Tumor burden was measured by bioluminescence.ResultsWe demonstrated that a fourth- generation CD19-targeted CAR (CAR.19) co-expressing IL-15 linked to its receptor IL-15/IL-15Rα (CAR.19-IL-15/IL-15Rα) significantly enhanced NK-92 cell proliferation, proinflammatory cytokine secretion, and cytotoxic activity against B-cell cancer cell lines in vitro and in a xenograft mouse model.ConclusionTogether with the results of the systematic analysis of the transcriptome of activated NK-92 CAR variants, this supports the notion that IL-15/IL-15Rα comprising fourth-generation CARs may overcome the limitations of NK-92 cell-based targeted tumor therapies in vivo by providing the necessary growth and activation signals

Catálogo Taxonômico da Fauna do Brasil: setting the baseline knowledge on the animal diversity in Brazil

The limited temporal completeness and taxonomic accuracy of species lists, made available in a traditional manner in scientific publications, has always represented a problem. These lists are invariably limited to a few taxonomic groups and do not represent up-to-date knowledge of all species and classifications. In this context, the Brazilian megadiverse fauna is no exception, and the Catálogo Taxonômico da Fauna do Brasil (CTFB) (http://fauna.jbrj.gov.br/), made public in 2015, represents a database on biodiversity anchored on a list of valid and expertly recognized scientific names of animals in Brazil. The CTFB is updated in near real time by a team of more than 800 specialists. By January 1, 2024, the CTFB compiled 133,691 nominal species, with 125,138 that were considered valid. Most of the valid species were arthropods (82.3%, with more than 102,000 species) and chordates (7.69%, with over 11,000 species). These taxa were followed by a cluster composed of Mollusca (3,567 species), Platyhelminthes (2,292 species), Annelida (1,833 species), and Nematoda (1,447 species). All remaining groups had less than 1,000 species reported in Brazil, with Cnidaria (831 species), Porifera (628 species), Rotifera (606 species), and Bryozoa (520 species) representing those with more than 500 species. Analysis of the CTFB database can facilitate and direct efforts towards the discovery of new species in Brazil, but it is also fundamental in providing the best available list of valid nominal species to users, including those in science, health, conservation efforts, and any initiative involving animals. The importance of the CTFB is evidenced by the elevated number of citations in the scientific literature in diverse areas of biology, law, anthropology, education, forensic science, and veterinary science, among others

Multipotent mesenchymal stromal cells promote melanoma metastasis through activation of the epithelial-to-mesenchymal transition

A interação entre células tumorais e células estromais tem um papel central na progressão neoplásica. As células estromais mesenquimais multipotentes (MSCs) podem se integrar ao microambiente tumoral onde modulam o crescimento dos tumores por meio de distintos mecanismos. Entretanto, pouco se sabe sobre o papel das MSCs na metástase, a principal causa de morte em pacientes com câncer. Utilizando um modelo de melanoma murino ortotópico, nós demonstramos que MSCs obtidas da medula óssea de camundongos (MO-MSCs) ocupam o nicho perivascular nos tumores primários e aumentam 2,5 vezes a incidência de micrometástases pulmonares quando co-infundidas com células de melanoma B16. Observamos ainda que o meio condicionado das MO-MSCs não altera o potencial de colonização pulmonar das células B16 infundidas sistemicamente. Isto indica que as MO-MSCs modulam as fases iniciais da cascata metastática, durante a qual ocorrem os processos de invasão e intravasão nos vasos sangüíneos. Em correlação com estes efeitos pró-metastáticos, o secretoma das MO-MSCs induziu a transição epitélio-mesenquimal (EMT) nas células de melanoma in vitro. Após cultivo em meio condicionado das MO-MSCs, as células B16 adquiriram uma morfologia evidentemente fibroblástica. Ao mesmo tempo, houve o rearranjo dos filamentos de actina e o aumento da expressão de marcadores mesenquimais como fibronectina, vimentina, FSP1, N-caderina e ZEB2, acompanhado da repressão transcricional de E-caderina. A ativação da EMT pelo secretoma das MO-MSCs resultou na aquisição de propriedades metastáticas nas células de melanoma. Após cultivo em meio condicionado de MO-MSCs, as células B16 tiveram seu potencial de ancoragem à fibronectina reduzido, ao passo que houve o aumento na mobilidade e no potencial de invasão em matrizes tridimensionais. Utilizando inibidores competitivos de ATP contra o receptor tirosina-cinase Met, demonstramos que a aquisição de todas as propriedades metastáticas avaliadas e a ativação da EMT nas células de melanoma é mediada pela ativação da via HGF/Met. Estes dados destacam o papel das MOMSCs no microambiente tumoral como fonte perivascular de moléculas indutoras da EMT, cuja ativação leva a aquisição de traços metastáticos nas células de melanoma. Além disso, a inibição da via HGF/Met pode neutralizar os efeitos das MO-MSCs sobre as células tumorais, contribuindo para a repressão de propriedades fundamentais que sustentam a progressão e a disseminação neoplásica. Estas informações são importantes para o desenvolvimento seguro das MO-MSCs como ferramenta terapêutica e demonstram a importância da sinalização entre MSCs e células tumorais na disseminação metastática. Mais especificamente, estas observações reforçam a inibição da via HGF/Met como uma abordagem promissora para o tratamento da metástase.The crosstalk between tumor cells and stromal cells can profoundly impact tumor progression. Multipotent mesenchymal stromal cells (MSCs) have been reported to integrate the tumor microenvironment where they are described to modulate tumor growth by distinct mechanisms. However, little is known about the impact of MSCs on metastasis, the main cause of death in patients with cancer. Using an orthotopic mouse melanoma model, we showed that mouse bone marrow-derived MSCs (BMMSCs) occupy the perivascular niche within primary tumors and increased by 2.5-fold the incidence of lung micrometastases after co-infusion with B16 melanoma cells. Also, MO-MSCs conditioned medium did not affect the lung colonization ability of systemically infused B16 cells. This indicates that MO-MSCs induces the initial steps of the metastatic cascade, during which the invasion and intravasion occurs. Correlating with these metastatic effects, the BM-MSCs\' secretome activated the epithelial-to-mesenchymal transition (EMT) in B16 cells in vitro. After culture in BMMSCs\' conditioned medium, B16 cells acquired an evident fibroblastic morphology. Simultaneously, we observed the rearrangement of actin filaments and the upregulation of mesenchymal markers such as fibronectin, vimentin, FSP1, Ncadherin and ZEB2. In agreement with the loss of epithelial phenotype, BM-MSCs\' secretome also suppressed E-cadherin expression in B16 cells. The activation of EMT by BM-MSCs leaded to the acquisition of metastatic traits in melanoma cells. After culture in BM-MSCs\' conditioned medium, B16 cells displayed reduced anchorage to fibronectin and increased motility and invasiveness in threedimensional matrix plugs. Inhibition of Met receptor with competitive ATP inhibitors demonstrated that the induction of EMT and the resultant acquisition of metastatic traits are driven by activation of HGF/Met signaling pathway. Taken together, these evidences highlight the role of BM-MSCs as a perivascular source of EMT-inductive signals, whose activation leads to acquisition of metastatic traits in melanoma cells. Furthermore, inhibition of HGF/Met signaling pathway can neutralize the effects of BM-MSCs on tumor cells, thereby allowing the repression of fundamental properties which support tumor progression and metastasis. This information is useful to safely develop BM-MSCs as therapeutic tool and demonstrate the relevance of the signaling between MSCs and tumor cells during metastasis. More specifically, it reinforces that inhibition of Met signaling can be a promissory approach for the treatment of metastasis

Generation of vascularized organoids as vehicles for production of coagulation factor VIII in a mouse model of hemophilia A

A hemofilia A (HA) é uma doença hemorrágica causada por mutações que levam à deficiência na produção ou função do fator VIII da coagulação (FVIII). O desafio atual é estabelecer uma terapia profilática e duradoura capaz de assegurar níveis plasmáticos de FVIII suficientes para corrigir o fenótipo da HA. Uma das estratégias é modificar células ex vivo para expressar o FVIII e transplantá-las para que secretem a referida proteína in vivo. No entanto, esta abordagem tem sido limitada pela sobrevida curta das células transplantadas. Neste trabalho, hipotetizamos que células endoteliais vasculares da veia do cordão umbilical (HUVECs) e células progenitoras perivasculares como as células estromais do tecido adiposo (ADSCs) poderiam ser co-transplantadas para formar organoides vascularizados que serviriam como veículos produtores de FVIII in vivo. Para isso, primeiramente investigamos o potencial do meio para crescimento de células endoteliais (EGM-2) contendo fatores de crescimento pró-angiogênicos na manutenção das propriedades progenitoras e pró-angiogênicas das ADSCs após cultivo. Para tanto, primeiramente comparamos o efeito de formulações de meios convencionais (DMEM + 10% de soro fetal bovino [DMEM] e αMEM + 15% de SFB [αMEM]) e do EGM-2 na eficiência clonogênica, proliferação e diferenciação in vitro das ADSCs. ADSCs cultivadas em EGM-2 (ADSC-EGM) apresentaram a maior eficiência clonogênica, maior potencial proliferativo e maior capacidade de diferenciação adipogênica e osteogênica, seguidas pelas ADSC-αMEM e pelas ADSC-DMEM, respectivamente. Em seguida, investigamos o potencial das ADSC-EGM e ADSC-αMEM em dar suporte à sobrevida das HUVECs e de estimular a angiogênese. Para isso, modificamos HUVECs para expressar a proteína repórter bioluminescente luciferase 2 e utilizamos o sinal bioluminescente para monitorar a sobrevida das mesmas. Após co-cultivo em meio reduzido de fatores de crescimento, apenas as ADSC-EGM atenuaram a apoptose das HUVECluc, sendo que a sobrevida das HUVECluc foi diretamente proporcional à quantidade de ADSC-EGM nos co-cultivos. Além disso, apenas as ADSC-EGM estimularam a formação de cordões endoteliais pelas HUVECluc durante o co-cultivo sob privação de fatores de crescimento. Notadamente, a extensão e ramificação da rede de cordões endoteliais formada foi diretamente proporcional à quantidade de ADSC-EGM. Estes efeitos anti-apoptótico e pró-angiogênico das ADSC-EGM foram recapitulados quando utilizamos apenas o meio condicionado das ADSC-EGM, indicando que o secretoma das ADSCs é alterado após cultivo em EGM-2. Em seguida, ADSCs e HUVECluc foram co-transplantadas subcutaneamente em camundongos imunodeficientes e a sobrevida das HUVECluc foi avaliada pela quantificação da bioluminescência. Após 60 dias, a quantidade de HUVECluc vivas foi equivalente entre aquelas transplantadas sozinhas ou com ADSC-αMEM (15,0%±0,7% vs 13,0%±0,5%, respectivamente). Entretanto, a sobrevida das HUVECluc co-transplantadas com ADSCEGM atingiu 105%±3,5%. Análises histológicas demonstraram que os organoides gerados pela co-infusão de HUVECluc e ADSC-EGM apresentaram maior densidade vascular e os vasos sanguíneos eram funcionais, a julgar pela presença de eritrócitos em seus lúmens. Em seguida, as ADSC-EGM e HUVECs foram modificadas para expressar o FVIII humano com o domínio B parcialmente deletado e transplantadas subcutaneamente em camundongos hemofílicos A imunodeficientes, os quais foram gerados após o cruzamento de machos NSG com fêmeas hemofílicas. Em todas as tentativas de transplante não houve aumento na atividade biológica do FVIII no plasma dos animais. Para avaliar se a rota de infusão estava influenciando a distribuição do FVIII na circulação, transplantamos células de adenocarcinoma hepático SK-HEP expressando FVIII e luciferase pelas vias subcutânea e intra-hepática. Após 8 dias, apenas os animais que receberam o transplante intra-hepático apresentaram uma atividade biológica do FVIII no plasma superior a 0,5 UI/mL. Após 15 dias, a atividade detectada foi pelo menos de 1 UI/mL, ao passo que os animais transplantados subcutaneamente não apresentavam níveis detectáveis de FVIII no plasma. Utilizando dados de bioluminescência, observamos que as células transplantadas subcutaneamente ou no fígado apresentavam a mesma velocidade de crescimento. Em suma, nosso dados demonstram que o cultivo das ADSCs em um microambiente pró-angiogênico aumenta sua eficiência clonogênica, sua capacidade de diferenciação in vitro e seu potencial em promover a sobrevida de células endoteliais e estimular a angiogênese in vivo em comparação com estratégias de cultivo convencionais. Estas observações abrem caminho para a identificação de moléculas importantes na regulação da multipotência e da atividade pró-angiogênica das ADSCs. Além disso, nossos achados estabelecem uma estratégia capaz de aumentar a eficiência terapêutica das ADSCs contra doenças isquêmicas e de elevar a sobrevida de células endoteliais co-transplantadas para servir como veículo produtor de proteínas terapêuticas in vivo. Por fim, concluímos que o local de implante das células expressando FVIII é crítico para a eficiência terapêutica deste tipo de terapia celular para a correção da HA.Hemophilia A (HA) is a hemorrhagic disorder caused by a deffective production or function of the coagulation factor VIII (FVIII). One of the frontiers in HA treatment is to establish a long-lasting prophylactic therapy to provide sufficient plasma levels of FVIII to correct the bleeding phenotype. One of the strategies to accomplish this is to modify cells ex vivo to express FVIII and transplant them to secrete the therapeutic protein in vivo. However, this approach has been limited by the poor survival of the transplanted cells. In this work, we hypothesized that the human umbilical vein endothelial cells (HUVECs) and perivascular progenitor cells such as the adipose-derived stromal cells (ADSCs) could be co-transplanted to form vascularized organoids which could serve as vehicles for FVIII production in vivo. For this, we innitially investigated the potential of endothelial cell growth medium (EGM-2) containing pro-angiogenic growth factors on the maintenance of progenitor and pro-angiogenic properties of ADSCs after culture. To this end, we first compared the effect of conventional media formulations (DMEM + 10% fetal bovine serum [DMEM], and αMEM + 15% FBS [αMEM]) and EGM-2 on the clonogenic efficiency, proliferation and in vitro differentiation of ADSCs. ADSCs cultured in EGM-2 (ADSC-EGM) presented the highest clonogenic efficiency, proliferative potential and adipogenic and osteogenic differentiation abilities, followed by the ADSCs cultured in αMEM (ADSC-αMEM) and in DMEM (ADSC-DMEM), respectively. Next, we investigated the potential of ADSC-EGM and ADSC-αMEM to support the survival of HUVECs and to promote angiogenesis. For this purpose, HUVECs were modified to express the bioluminescent enzime luciferase 2 and the bioluminescent signal was used to monitor cell survival. After co-culture in a growth factor reduced medium (i.e. αMEM+1% FBS), only the ADSC-EGM attenuated HUVECluc apoptosis, and the survival of HUVECluc was directly proportional to the amount of ADSC-EGM seeded. Furthermore, only the ADSCEGM were able to stimulate the formation of endothelial cords by HUVECluc during coculture under growth factor starvation. Notably, the length and branching of the endothelial cords were directly proportional to the amount of ADSC-EGM in the cocultures. These anti-apoptotic and pro-angiogenic effects of ADSC-EGM were recapitulated by the ADSC-EGM conditioned medium, indicating that the secretome of the ADSCs is altered upon culture in EGM-2. Next, HUVECluc and ADSCs were cotransplanted subcutaneously into immunodeficient mice and the survival of HUVECluc was monitored by bioluminescent imaging. After 60 days the amount of living HUVECluc was equivalent between those transplanted with HUVECluc alone and those co-transplanted with HUVECluc and ADSC-αMEM (15.0%±0.7% vs. 13.0%±0.5%, respectively). However, the survival of HUVECluc co-transplanted with ADSC-EGM reached 105.0%±3.5%. Histological analysis showed that the organoids generated by the co-transplanation of HUVECluc and ADSC-EGM had the higher vascular density and that the blood vessels formed were functional, as indicated by the presence of erythrocytes within their lumens. Next, the ADSC-EGM and HUVECs were engineered to express human FVIII with the Bdomain partially deleted and were transplanted subcutaneously in immunodeficient hemophilic A mice, which were generated after crossing NSG males with hemophilic A females. All attempts of transplantation did not resulted in any increase of the biological activity of FVIII in the plasma of the animals. To evaluate whether the site of transplantation was influencing the distribution of FVIII to the circulation, we transplanted liver adenocarcinoma SK-HEP cells expressing both FVIII and luciferase 2 by subcutaneous and intrahepatic injections. After 8 days, the animals receiving the intrahepatic injections displayed plasma levels of FVIII above 0.5 IU/mL. After 15 days, the FVIII levels increased to at least 1.0 IU/mL. In a sharp contrast, animals that received subcutaneous injections showed no increase in the plasmatic levels of FVIII. Using bioluminescence quantification, we observed that the cells transplanted subcutaneously or intrahepatically showed similar growth rates in vivo. Collectively, these findings demonstrate that the cultivation of the ADSCs in a pro-angiogenic millieu increases their clonogenic efficiency, in vitro differentiation capacity and their potential to promote survival of endothelial cells and stimulate angiogenesis in vivo. These findings lay the groundwork for identifying key modulators of ADSCs multipotency and proangiogenic properties. Moreover, our pre-conditioning strategy may be explored as a strategy to increase the therapeutic efficiency of ADSCs against ischemic diseases and to increase the survival of co-transplanted endothelial cells to serve as a vehicle for delivering therapeutic proteins in vivo. Finally, we conclude that the implantation site of FVIII-expressing cells is crucial to achieve the correction of HA

Multipotent mesenchymal stromal cells promote melanoma metastasis through activation of the epithelial-to-mesenchymal transition

A interação entre células tumorais e células estromais tem um papel central na progressão neoplásica. As células estromais mesenquimais multipotentes (MSCs) podem se integrar ao microambiente tumoral onde modulam o crescimento dos tumores por meio de distintos mecanismos. Entretanto, pouco se sabe sobre o papel das MSCs na metástase, a principal causa de morte em pacientes com câncer. Utilizando um modelo de melanoma murino ortotópico, nós demonstramos que MSCs obtidas da medula óssea de camundongos (MO-MSCs) ocupam o nicho perivascular nos tumores primários e aumentam 2,5 vezes a incidência de micrometástases pulmonares quando co-infundidas com células de melanoma B16. Observamos ainda que o meio condicionado das MO-MSCs não altera o potencial de colonização pulmonar das células B16 infundidas sistemicamente. Isto indica que as MO-MSCs modulam as fases iniciais da cascata metastática, durante a qual ocorrem os processos de invasão e intravasão nos vasos sangüíneos. Em correlação com estes efeitos pró-metastáticos, o secretoma das MO-MSCs induziu a transição epitélio-mesenquimal (EMT) nas células de melanoma in vitro. Após cultivo em meio condicionado das MO-MSCs, as células B16 adquiriram uma morfologia evidentemente fibroblástica. Ao mesmo tempo, houve o rearranjo dos filamentos de actina e o aumento da expressão de marcadores mesenquimais como fibronectina, vimentina, FSP1, N-caderina e ZEB2, acompanhado da repressão transcricional de E-caderina. A ativação da EMT pelo secretoma das MO-MSCs resultou na aquisição de propriedades metastáticas nas células de melanoma. Após cultivo em meio condicionado de MO-MSCs, as células B16 tiveram seu potencial de ancoragem à fibronectina reduzido, ao passo que houve o aumento na mobilidade e no potencial de invasão em matrizes tridimensionais. Utilizando inibidores competitivos de ATP contra o receptor tirosina-cinase Met, demonstramos que a aquisição de todas as propriedades metastáticas avaliadas e a ativação da EMT nas células de melanoma é mediada pela ativação da via HGF/Met. Estes dados destacam o papel das MOMSCs no microambiente tumoral como fonte perivascular de moléculas indutoras da EMT, cuja ativação leva a aquisição de traços metastáticos nas células de melanoma. Além disso, a inibição da via HGF/Met pode neutralizar os efeitos das MO-MSCs sobre as células tumorais, contribuindo para a repressão de propriedades fundamentais que sustentam a progressão e a disseminação neoplásica. Estas informações são importantes para o desenvolvimento seguro das MO-MSCs como ferramenta terapêutica e demonstram a importância da sinalização entre MSCs e células tumorais na disseminação metastática. Mais especificamente, estas observações reforçam a inibição da via HGF/Met como uma abordagem promissora para o tratamento da metástase.The crosstalk between tumor cells and stromal cells can profoundly impact tumor progression. Multipotent mesenchymal stromal cells (MSCs) have been reported to integrate the tumor microenvironment where they are described to modulate tumor growth by distinct mechanisms. However, little is known about the impact of MSCs on metastasis, the main cause of death in patients with cancer. Using an orthotopic mouse melanoma model, we showed that mouse bone marrow-derived MSCs (BMMSCs) occupy the perivascular niche within primary tumors and increased by 2.5-fold the incidence of lung micrometastases after co-infusion with B16 melanoma cells. Also, MO-MSCs conditioned medium did not affect the lung colonization ability of systemically infused B16 cells. This indicates that MO-MSCs induces the initial steps of the metastatic cascade, during which the invasion and intravasion occurs. Correlating with these metastatic effects, the BM-MSCs\' secretome activated the epithelial-to-mesenchymal transition (EMT) in B16 cells in vitro. After culture in BMMSCs\' conditioned medium, B16 cells acquired an evident fibroblastic morphology. Simultaneously, we observed the rearrangement of actin filaments and the upregulation of mesenchymal markers such as fibronectin, vimentin, FSP1, Ncadherin and ZEB2. In agreement with the loss of epithelial phenotype, BM-MSCs\' secretome also suppressed E-cadherin expression in B16 cells. The activation of EMT by BM-MSCs leaded to the acquisition of metastatic traits in melanoma cells. After culture in BM-MSCs\' conditioned medium, B16 cells displayed reduced anchorage to fibronectin and increased motility and invasiveness in threedimensional matrix plugs. Inhibition of Met receptor with competitive ATP inhibitors demonstrated that the induction of EMT and the resultant acquisition of metastatic traits are driven by activation of HGF/Met signaling pathway. Taken together, these evidences highlight the role of BM-MSCs as a perivascular source of EMT-inductive signals, whose activation leads to acquisition of metastatic traits in melanoma cells. Furthermore, inhibition of HGF/Met signaling pathway can neutralize the effects of BM-MSCs on tumor cells, thereby allowing the repression of fundamental properties which support tumor progression and metastasis. This information is useful to safely develop BM-MSCs as therapeutic tool and demonstrate the relevance of the signaling between MSCs and tumor cells during metastasis. More specifically, it reinforces that inhibition of Met signaling can be a promissory approach for the treatment of metastasis

BMAL1 Knockdown Leans Epithelial–Mesenchymal Balance toward Epithelial Properties and Decreases the Chemoresistance of Colon Carcinoma Cells

The circadian clock coordinates biological and physiological functions to day/night cycles. The perturbation of the circadian clock increases cancer risk and affects cancer progression. Here, we studied how BMAL1 knockdown (BMAL1-KD) by shRNA affects the epithelial–mesenchymal transition (EMT), a critical early event in the invasion and metastasis of colorectal carcinoma (CRC). In corresponding to a gene set enrichment analysis, which showed a significant enrichment of EMT and invasive signatures in BMAL1_high CRC patients as compared to BMAL1_low CRC patients, our results revealed that BMAL1 is implicated in keeping the epithelial–mesenchymal equilibrium of CRC cells and influences their capacity of adhesion, migration, invasion, and chemoresistance. Firstly, BMAL1-KD increased the expression of epithelial markers (E-cadherin, CK-20, and EpCAM) but decreased the expression of Twist and mesenchymal markers (N-cadherin and vimentin) in CRC cell lines. Finally, the molecular alterations after BMAL1-KD promoted mesenchymal-to-epithelial transition-like changes mostly appeared in two primary CRC cell lines (i.e., HCT116 and SW480) compared to the metastatic cell line SW620. As a consequence, migration/invasion and drug resistance capacities decreased in HCT116 and SW480 BMAL1-KD cells. Together, BMAL1-KD alerts the delicate equilibrium between epithelial and mesenchymal properties of CRC cell lines, which revealed the crucial role of BMAL1 in EMT-related CRC metastasis and chemoresistance

Sex and Circadian Timing Modulate Oxaliplatin Hematological and Hematopoietic Toxicities

Oxaliplatin was nearly twice as hematotoxic, with optimal circadian timing differing by 6 h, in women as compared to men with colorectal cancers. Hence, we investigated sex- and timing-related determinants of oxaliplatin hematopoietic toxicities in mice. Body-weight loss (BWL), blood cell counts, bone marrow cellularity (BMC) and seven flow-cytometry-monitored hematopoietic progenitor populations were evaluated 72 h after oxaliplatin chronotherapy administration (5 mg/kg). In control animals, circadian rhythms of circulating white blood cells showed a peak at ZT5 in both sexes, whereas BMC was maximum at ZT20 in males and ZT13h40 in females. All BM progenitor counts presented robust rhythms with phases around ZT3h30 in females, whereas only three of them rhythmically cycled in males with a ≈ −6 h phase shift. In treated females, chronotoxicity rhythms occurred in BWL, WBC, BMC and all BM progenitors with the best timing at ZT15, ZT21, ZT15h15 and ZT14h45, respectively. In males, almost no endpoints showed circadian rhythms, BWL and WBC toxicity being minimal, albeit with a substantial drop in BM progenitors. Increasing dose (10 mg/kg) in males induced circadian rhythms in BWL and WBC but not in BM endpoints. Our results suggest complex and sex-specific clock-controlled regulation of the hematopoietic system and its response to oxaliplatin