RESUMO
BACKGROUND: Enhanced recovery after surgery (ERAS) protocols have been shown to reduce length of stay (LOS) and complications. The impact of ERAS protocols on the cost of cytoreductive surgery and hyperthermic intraperitoneal chemotherapy (CRS-HIPEC) has not been studied. PATIENTS AND METHODS: We performed a retrospective cohort analysis of patients undergoing CRS-HIPEC from 2016-2022 at a single quaternary center. Propensity score matching was used to create pre-and post-ERAS cohorts. Cost, overall and serious complications, and intensive care unit (ICU) length of stay (LOS) between the two cohorts were compared using the Mann-Whitney U-test for continuous variables and χ2 test for categorical variables. RESULTS: Our final matched cohort consisted of 100 patients, with 50 patients in both the pre- and post-ERAS groups. After adjusting for patient complexity and inflation, the median total cost [$75,932 ($67,166-102,645) versus $92,992 ($80,720-116,710), p = 0.02] and operating room cost [$26,817 ($23,378-33,121) versus $34,434 ($28,085-$41,379), p < 0.001] were significantly higher in the post-ERAS cohort. Overall morbidity (n = 22, 44% versus n = 17, 34%, p = 0.40) and ICU length of stay [2 days (IQR 1-3) versus 2 days (IQR 1-4), p = 0.70] were similar between the two cohorts. A total cost increase of $22,393 [SE $13,047, 95% CI (-$3178 to $47,965), p = 0.086] was estimated after implementation of ERAS, with operating room cost significantly contributing to this increase [$8419, SE $1628, 95% CI ($5228-11,609), p < 0.001]. CONCLUSIONS: CRS-HIPEC ERAS protocols were associated with higher total costs due to increased operating room costs at a single institution. There was no significant difference in ICU LOS and complications after the implementation of the ERAS protocol.
Assuntos
Procedimentos Cirúrgicos de Citorredução , Recuperação Pós-Cirúrgica Melhorada , Quimioterapia Intraperitoneal Hipertérmica , Tempo de Internação , Neoplasias Peritoneais , Humanos , Feminino , Procedimentos Cirúrgicos de Citorredução/economia , Masculino , Estudos Retrospectivos , Quimioterapia Intraperitoneal Hipertérmica/economia , Pessoa de Meia-Idade , Tempo de Internação/estatística & dados numéricos , Neoplasias Peritoneais/terapia , Terapia Combinada , Seguimentos , Complicações Pós-Operatórias , Prognóstico , Idoso , Unidades de Terapia Intensiva/economia , Quimioterapia do Câncer por Perfusão Regional/economia , Taxa de SobrevidaRESUMO
The disruption of ordered water molecules confined within hydrophobic reaction pockets alters the energetics of adsorption and catalysis, but a mechanistic understanding of how nonaqueous solvents influence catalysis in microporous voids remains unclear. Here, we use kinetic analyses coupled with IR spectroscopy to study how alkanol hydrogen-bonding networks confined within hydrophobic and hydrophilic zeolite catalysts modify reaction free energy landscapes. Hydrophobic Beta zeolites containing framework Sn atoms catalyze the transfer hydrogenation reaction of cyclohexanone in a 2-butanol solvent 10× faster than their hydrophilic analogues. This rate enhancement stems from the ability of hydrophobic Sn-Beta to inhibit the formation of extended liquid-like 2-butanol oligomers and promote dimeric H-bonded 2-butanol networks. These different intraporous 2-butanol solvent structures manifest as differences in the activation and adsorption enthalpies and entropies that comprise the free energy landscape of transfer hydrogenation catalysis. The ordered H-bonding solvent network present in hydrophobic Sn-Beta stabilizes the transfer hydrogenation transition state to a greater extent than the liquid-like 2-butanol solvent present in hydrophilic Sn-Beta, giving rise to higher turnover rates on hydrophobic Sn-Beta. Additionally, reactant adsorption within hydrophobic Sn-Beta is driven by the breakup of intraporous solvent-solvent interactions, resulting in positive enthalpies of adsorption that are partially compensated by an increase in the solvent reorganization entropy. Collectively, these results emphasize the ability of the zeolite pore to regulate the structure of confined nonaqueous H-bonding solvent networks, which offers an additional dimension to modulate adsorption and reactivity.
Assuntos
Procedimentos Cirúrgicos de Citorredução , Quimioterapia Intraperitoneal Hipertérmica , Humanos , Procedimentos Cirúrgicos de Citorredução/economia , Procedimentos Cirúrgicos de Citorredução/métodos , Terapia Combinada , Neoplasias Peritoneais/terapia , Recuperação Pós-Cirúrgica Melhorada , PrognósticoRESUMO
Inflammation-associated fibroblasts (IAFs) are associated with progression and drug resistance of chronic inflammatory diseases such as inflammatory bowel disease (IBD), but their direct impact on epithelial cells is unknown. Here, we developed an in vitro model whereby human colon fibroblasts are induced by specific cytokines and recapitulate key features of IAFs in vivo. When cocultured with patient-derived colon organoids (colonoids), IAFs induced rapid colonoid expansion and barrier disruption due to swelling and rupture of individual epithelial cells. Colonoids cocultured with IAFs also show increased DNA damage, mitotic errors, and proliferation arrest. These IAF-induced epithelial defects are mediated by a paracrine pathway involving prostaglandin E2 and its receptor EP4, leading to protein kinase A -dependent activation of the cystic fibrosis transmembrane conductance regulator. EP4-specific chemical inhibitors effectively prevented IAF-induced colonoid swelling and restored normal proliferation and genome stability. These findings reveal a mechanism by which IAFs could promote and perpetuate IBD and suggest a therapeutic avenue to mitigate inflammation-associated epithelial injury.
Assuntos
Doenças Inflamatórias Intestinais , Prostaglandinas , Humanos , Epitélio/metabolismo , Inflamação , Doenças Inflamatórias Intestinais/etiologia , Doenças Inflamatórias Intestinais/metabolismo , Fibroblastos/metabolismoRESUMO
Aneuploidy is nearly ubiquitous in tumor genomes, but the role of aneuploidy in the early stages of cancer evolution remains unclear. Here, by inducing heterogeneous aneuploidy in non-transformed human colon organoids (colonoids), we investigated how the effects of aneuploidy on cell growth and differentiation may promote malignant transformation. Previous work implicated p53 activation as a downstream response to aneuploidy induction. We found that simple aneuploidy, characterized by 1-3 gained or lost chromosomes, resulted in little or modest p53 activation and cell cycle arrest when compared with more complex aneuploid cells. Single-cell RNA sequencing analysis revealed that the degree of p53 activation was strongly correlated with karyotype complexity. Single-cell tracking showed that cells could continue to divide despite the observation of one to a few lagging chromosomes. Unexpectedly, colonoids with simple aneuploidy exhibited impaired differentiation after niche factor withdrawal. These findings demonstrate that simple aneuploid cells can escape p53 surveillance and may contribute to niche factor-independent growth of cancer-initiating colon stem cells.
Assuntos
Aneuploidia , Diferenciação Celular , Proliferação de Células , Organoides , Proteína Supressora de Tumor p53 , Humanos , Proteína Supressora de Tumor p53/metabolismo , Proteína Supressora de Tumor p53/genética , Organoides/metabolismo , Colo/metabolismo , Intestinos , Células-Tronco Neoplásicas/metabolismo , Células-Tronco Neoplásicas/patologia , Pontos de Checagem do Ciclo Celular/genética , Transformação Celular Neoplásica/genéticaRESUMO
T cell activation involves a cascade of TCR-mediated signals that are regulated by three distinct intracellular signaling motifs located within the cytoplasmic tails of the CD3 chains. Whereas all the CD3 subunits possess at least one ITAM, the CD3 ε subunit also contains a proline-rich sequence and a basic-rich stretch (BRS). The CD3 ε BRS complexes selected phosphoinositides, interactions that are required for normal cell surface expression of the TCR. The cytoplasmic domain of CD3 ζ also contains several clusters of arginine and lysine residues. In this study, we report that these basic amino acids enable CD3 ζ to complex the phosphoinositides PtdIns(3)P, PtdIns(4)P, PtdIns(5)P, PtdIns(3,5)P(2), and PtdIns(3,4,5)P(3) with high affinity. Early TCR signaling pathways were unaffected by the targeted loss of the phosphoinositide-binding functions of CD3 ζ. Instead, the elimination of the phosphoinositide-binding function of CD3 ζ significantly impaired the ability of this invariant chain to accumulate stably at the immunological synapse during T cell-APC interactions. Without its phosphoinositide-binding functions, CD3 ζ was concentrated in intracellular structures after T cell activation. Such findings demonstrate a novel functional role for CD3 ζ BRS-phosphoinositide interactions in supporting T cell activation.
Assuntos
Complexo CD3/metabolismo , Sinapses Imunológicas , Fosfatidilinositóis/metabolismo , Complexo Receptor-CD3 de Antígeno de Linfócitos T/metabolismo , Aminoácidos Básicos , Animais , Sítios de Ligação/imunologia , Complexo CD3/química , Complexo CD3/imunologia , Linhagem Celular , Humanos , Ativação Linfocitária/imunologia , Camundongos , Fosfatidilinositóis/imunologia , Ligação Proteica/imunologia , Transdução de Sinais/imunologia , TransfecçãoRESUMO
Inflammation-associated fibroblasts (IAFs) are associated with the progression and drug resistance of chronic inflammatory diseases such as inflammatory bowel disease (IBD), but their direct impact on epithelial function and architecture is unknown. In this study, we developed an in vitro model whereby human colon fibroblasts are induced to become IAFs by specific cytokines and recapitulate key features of IAFs in vivo. When co-cultured with patient-derived colon organoids (colonoids), IAFs induced rapid colonoid swelling and barrier disruption due to swelling and rupture of individual epithelial cells. Epithelial cells co-cultured with IAFs also exhibit increased DNA damage, mitotic errors, and proliferation arrest. These IAF-induced epithelial defects are mediated through a paracrine pathway involving prostaglandin E2 (PGE2) and the PGE2 receptor EP4, leading to PKA-dependent activation of the CFTR chloride channel. Importantly, EP4-specific chemical inhibitors effectively prevented colonoid swelling and restored normal proliferation and genome stability of IAF-exposed epithelial cells. These findings reveal a mechanism by which IAFs could promote and perpetuate IBD and suggest a potential treatment to mitigate inflammation-associated epithelial injury.
RESUMO
Aneuploidy, a near ubiquitous genetic feature of tumors, is a context-dependent driver of cancer evolution; however, the mechanistic basis of this role remains unclear. Here, by inducing heterogeneous aneuploidy in non-transformed human colon organoids (colonoids), we investigate how the effects of aneuploidy on cell growth and differentiation may promote malignant transformation. Single-cell RNA sequencing reveals that the gene expression signature across over 100 unique aneuploid karyotypes is enriched with p53 responsive genes. The primary driver of p53 activation is karyotype complexity. Complex aneuploid cells with multiple unbalanced chromosomes activate p53 and undergo G1 cell-cycle arrest, independent of DNA damage and without evidence of senescence. By contrast, simple aneuploid cells with 1-3 chromosomes gained or lost continue to proliferate, demonstrated by single cell tracking in colonoids. Notably, simple aneuploid cells exhibit impaired differentiation when niche factors are withdrawn. These findings suggest that while complex aneuploid cells are eliminated from the normal epithelium due to p53 activation, simple aneuploid cells can escape this checkpoint and may contribute to niche factor-independent growth of cancer-initiating cells.
RESUMO
TP53-dependent cell cycle arrest has been proposed to limit the proliferation of aneuploid cells. We investigated the cellular response to aneuploidy in cell lines and organoid cultures and found that TP53 (also known as p53) is not activated following aneuploidy induction in organoids. However, we confirmed that p53 is required for high mitotic fidelity. Our findings provide a revised view on how p53 safeguards against aneuploidy.
RESUMO
Although much is known about the gene mutations required to drive colorectal cancer (CRC) initiation, the tissue-specific selective microenvironments in which neoplasia arises remains less characterized. Here, we determined whether modulation of intestinal stem cell niche morphogens alone can exert a neoplasia-relevant selective pressure on normal colonic epithelium. Using adult stem cell-derived murine colonic epithelial organoids (colonoids), we employed a strategy of sustained withdrawal of epidermal growth factor (EGF) and epidermal growth factor receptor (EGFR) inhibition to select for and expand survivors. EGFR-signaling-independent (iEGFR) colonoids emerged over rounds of selection and expansion. Colonoids derived from a mouse model of chronic mucosal injury showed an enhanced ability to adapt to EGFR inhibition. Whole-exome and transcriptomic analyses of iEGFR colonoids demonstrated acquisition of deleterious mutations and altered expression of genes implicated in EGF signaling, pyroptosis, and CRC. iEGFR colonoids acquired dysplasia-associated cytomorphologic changes, an increased proliferative rate, and the ability to survive independently of other required niche factors. These changes were accompanied by emergence of aneuploidy and chromosomal instability; further, the observed mitotic segregation errors were significantly associated with loss of interkinetic nuclear migration, a fundamental and dynamic process underlying intestinal epithelial homeostasis. This study provides key evidence that chromosomal instability and other phenotypes associated with neoplasia can be induced ex vivo via adaptation to EGF withdrawal in normal and stably euploid colonic epithelium, without introducing cancer-associated driver mutations. In addition, prior mucosal injury accelerates this evolutionary process.
Assuntos
Instabilidade Cromossômica , Colo/metabolismo , Mucosa Intestinal/metabolismo , Adaptação Biológica , Aneuploidia , Animais , Proliferação de Células , Células Cultivadas , Colo/patologia , Receptores ErbB/genética , Receptores ErbB/metabolismo , Edição de Genes , Regulação da Expressão Gênica , Genes APC , Humanos , Mucosa Intestinal/patologia , Camundongos , Mutação , Organoides , Técnicas de Cultura de TecidosRESUMO
Most solid tumors are aneuploid, and p53 has been implicated as the guardian of the euploid genome. Previous experiments using human cell lines showed that aneuploidy induction leads to p53 accumulation and p21-mediated G1 cell cycle arrest. We find that adherent 2-dimensional (2D) cultures of human immortalized or cancer cell lines activate p53 upon aneuploidy induction, whereas suspension cultures of a human lymphoid cell line undergo a p53-independent cell cycle arrest. Surprisingly, 3D human and mouse organotypic cultures from neural, intestinal, or mammary epithelial tissues do not activate p53 or arrest in G1 following aneuploidy induction. p53-deficient colon organoids have increased aneuploidy and frequent lagging chromosomes and multipolar spindles during mitosis. These data suggest that p53 may not act as a universal surveillance factor restricting the proliferation of aneuploid cells but instead helps directly or indirectly ensure faithful chromosome transmission likely by preventing polyploidization and influencing spindle mechanics.
Assuntos
Aneuploidia , Proteína Supressora de Tumor p53/metabolismo , Animais , Adesão Celular , Pontos de Checagem do Ciclo Celular , Proteínas de Ciclo Celular/antagonistas & inibidores , Proteínas de Ciclo Celular/metabolismo , Linhagem Celular , Colo/metabolismo , Humanos , Mamíferos , Camundongos Endogâmicos C57BL , Mitose , Organoides/metabolismo , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Tirosina Quinases/antagonistas & inibidores , Proteínas Tirosina Quinases/metabolismoRESUMO
Postsurgical spine imaging actually commences with the preoperative and perioperative imaging examinations that are performed before and during the patient's surgical procedure, respectively. It is in this context that postsurgical spine imaging examinations are best evaluated with a better appreciation of the changes, sometimes dramatic, that can occur following spine surgery. Careful follow-up is the rule in spine surgery, and these patients will have multiple imaging examinations over time. Some studies are performed immediately due to possible complications or exacerbation of pain symptoms, whereas others are used to assess the status of a fusion procedure. In any case it is prudent to always be aware of the overlap between the "normal" or expected imaging findings in the postoperative spine and potential pathologic processes that are developing at the postsurgical site. Additionally, a basic understanding of common spine surgical techniques and approaches is paramount toward rendering a thoughtful analysis. In this chapter, the authors discuss these imaging findings in the setting of the most commonly performed spine surgeries and emphasize the importance of active communication between the radiologist and spine surgeon.