Chemotherapeutic Injury Research Articles

Metabolite-based inter-kingdom communication controls intestinal tissue recovery following chemotherapeutic injury

Cytotoxic chemotherapies have devastating side effects, particularly within the gastrointestinal tract. Gastrointestinal toxicity includes the death and damage of the epithelium and an imbalance in the intestinal microbiota, otherwise known as dysbiosis. Whether dysbiosis is a direct contributor to tissue toxicity is a key area of focus. Here, from both mammalian and bacterial perspectives, we uncover an intestinal epithelial cell death-Enterobacteriaceae signaling axis that fuels dysbiosis. Specifically, our data demonstrate that chemotherapy-induced epithelial cell apoptosis and the purine-containing metabolites released from dying cells drive the inter-kingdom transcriptional re-wiring of the Enterobacteriaceae, including fundamental shifts in bacterial respiration and promotion of purine utilization-dependent expansion, which in turn delays the recovery of the intestinal tract. Inhibition of epithelial cell death or restriction of the Enterobacteriaceae to homeostatic levels reverses dysbiosis and improves intestinal recovery. These findings suggest that supportive therapies that maintain homeostatic levels of Enterobacteriaceae may be useful in resolving intestinal disease.

Ganoderma lucidum Polysaccharide Peptide Alleviates Cyclophosphamide-Induced Male Reproductive Injury by Reducing Oxidative Stress and Apoptosis.

Chemotherapy is an important factor leading to male infertility. It is crucial to discover safe and effective treatments to prevent male reproductive injury caused by chemotherapy. The Ganoderma lucidum polysaccharide peptide (GLPP) has multiple pharmacological activities. The purpose of this study was to determine whether GLPP could protect the male sperm production from chemotherapeutic injury using a mouse model, with testicular damage induced by cyclophosphamide (CP). CP (50 mg/kg/day) was injected intraperitoneally into male ICR mice gavaged with different doses of GLPP at certain spermatogenic stages. The experimental results showed that GLPP alleviated the CP-induced reduction in reproductive organ coefficients and sperm parameters and reduced the morphological damage of testicular tissues in a dose-dependent manner. GLPP significantly improved the reproductive index, sperm-related parameters, sex hormone levels, and histological testis architecture at different spermatogenic stages. Furthermore, GLPP significantly increased superoxide dismutase (SOD), glutathione (GSH), catalase (CAT), Nrf2, and HO-1, and decreased malondialdehyde (MDA) and Keap-1 in the testicular tissue, indicating reduced oxidative stress. In addition, GLPP limited CP-induced apoptosis via a reduction in Bax expression and increase in Bcl-2 expression. This study suggests that GLPP plays a protective role in spermatogenesis by reducing chemotherapeutic injury and might be developed into drug for male patients receiving chemotherapy.

Salvianolic Acid B Ameliorated Chemotherapeutic Injury of Cardiac Myocytes through the Nrf2/ARE Signaling Pathway.

Cardiotoxicity has been corroborated to be the toxic influence of cisplatin (CDDP). Oxidative stress and cardiomyocyte apoptosis play a vital part in cardiotoxicity induced by CDDP. Salvianolic acid Salvianolic acid B (SalB) is a monomeric component of Salvia miltiorrhiza, which has antioxidant and anti-inflammatory influences. In this research, we explored the mechanism of SalB in cardiotoxicity induced by CDDP. 36 Wistar rats were separated into sham subgroup, CDDP (10 mg/kg) subgroup, CDDP (10 mg/kg) + SalB (1 μM) subgroup at random, CDDP (10 mg/kg) + SalB (5 μM) subgroup and CDDP (10 mg/kg) + SalB (10 μM) subgroup, Nicotinic Acid Riboside (NAR, 5 μM), with 6 rats in each subgroup. The cardiac function of rats in each subgroup was estimated by echocardiography, and hematoxylin-eosin (HE) staining and Masson staining corroborated the pathological changes of cardiac tissue. Biochemical kits were utilized for detecting the lactate dehydrogenase (LDH), creatine kinase (CK), interleukin-1β (IL-1β), IL-18, and caspase-1 concentrations in serum, superoxide dismutase (SOD), and malondialdehyde (MDA) in myocardial tissue, TdT-mediated dUTP Nick-End Labeling (TUNEL) staining, and flow cytometry were utilized for estimating the apoptosis level in myocardial tissue, western blot was used for estimating caspase-3, Bcl2-Associated X (Bax) levels in myocardial tissue and proteins levels related to Nuclear factor E2 related factor 2 (Nrf2) signal pathway. CDDP-induced cardiac dysfunction, myocardial injury, boosted LDH and CK levels in serum (p < 0.05), memorably increased oxidative stress level in myocardial tissue (p < 0.05), boosted inflammatory response (p < 0.05), boosted apoptosis rate of cardiomyocytes (p < 0.05), and declined the Nrf2, NAD(P)H quinone oxidoreductase 1 (NQO1), heme oxygenase 1 (HO-1) protein levels (p < 0.05). Interestingly, SalB remedy could alleviate the changes caused by CDDP in the above parameters, significantly decrease the level of myocardial oxidative stress and apoptosis (p < 0.05). SalB ameliorates the injury of cardiomyocytes induced by chemotherapy through oxidative stress mediated by the Nrf2/antioxidant response element (ARE) signal pathway.

B4GALT1-Dependent Glycosylation Is Essential to Bone Marrow Reconstitution Following Chemotherapeutic Injury

The bone marrow (BM) comprises a three-dimensional (3D) network of branching sinusoids surrounding hematopoietic cells, embedded in an extracellular matrix (ECM) rich in glycans vital for cell maintenance and function. While the interaction between megakaryocytes (MKs) and the components of the environment for BM recovery post-injury becomes more apparent, the role of glycan-dependent thrombopoiesis and nascent platelets in BM maintenance and reconstitution is underexplored. We have found that β-1,4-galactosyltransferase 1 (β4galt1) deficiency results in dysplastic MKs, disrupted proplatelet release, and severe thrombocytopenia. KEGG analysis of SLAM marker sorted hematopoietic stem and progenitor cells (HSPC) bulk RNAseq data, showed downregulation of ECM-receptor interaction, focal adhesion, gap junction, and proteoglycans in cancer pathways in β4galt1 -/- HSPCs compared to controls. We hypothesized that β4galt1-dependent glycosylation regulates the BM microenvironment at steady state and post-injury. To test this, we conducted a study examining the effects of a single dose of 5-fluorouracil (5-FU) treatment (150 µg/g) on BM MKs, c-kit + progenitors, vasculature, ECM, and platelet count in both control and β4GalT1-deficient (B4 -/-) mice. We analyzed femur BM sections using immunofluorescence, targeting the vasculature marker laminin, progenitor marker c-kit, and ECM markers perlecan and fibrinogen, and MK marker GPIbα (CD42b) and CD41. Image processing with Imaris software and MATLAB facilitated cell count, spatial distribution, and 3D reconstruction assessments of the BM. An automated hemoanalyzer quantified platelet counts from days 1-10 post-treatment. Post-5-FU treatment, control mice exhibited a 50% decrease in reticulated and total platelet counts from day 3, persisting until day 8. On day 10, platelet counts increased to 150% above average. As BM cellularity diminished, a marrow sinusoidal cavity emerged at the bone's center, reaching maximum size on day 3 post-injury (mean size = 2.10 ± 0.14 µm 2; p=0.013), contracting from day 4 (mean size = 1.35 ± 0.14 µm 2; p=0.19), and restoring its size by day 8. GPIbα + MKs and c-kit + cell counts decreased by 50% 3 days following the injury while colocalization of c-kit + GPIbα + expressing cells increased preferentially localizing at the sinusoidal cavity, hinting at their role in BM recovery after injury. At day 5 post-injury, c-kit + cellcounts increased but reduced again on day 10, coinciding with the rise of MK number. 3D imaging post-5-FU treatment revealed a preferential localization of GPIbα + MKs and c-kit + cells in the center of the bone by day 10, adjacent to the previously injured area. High-resolution 3D reconstruction revealed that the sinusoidal cavity was filled with ECM components, including perlecan and fibrinogen, and platelets, as determined using GPIbα staining, on day 5 post-injury. Platelets were not released into circulation until the 8th day of reconstitution of BM, suggesting that although platelets are produced by BM MKs and distributed within the sinusoidal cavity, they do not immediately enter circulation. Depleting platelets using a platelet specific antibody before administering 5-FU slowed reconstitution of the BM sinusoidal cavity. In contrast to control mice, B4 -/- mice showed a reduction in platelet count and a notable accumulation of highly dysplastic MKs clusters around the sinusoidal cavity (MK/mm 3= 3598 vs. MK/mm 3=2748) at day 5 post-5FU. Moreover, SLAM marker defined long-term HSCs showed that MK-biased (CD41 + and CD41 +/ GPIbα +) populations notably increased in B4 -/- mice compared to controls (p=0.0043), indicating heightened emergency hematopoiesis in the absence of β4galt1. While the control BM sinusoidal cavity almost returned to normal size at day 8, the B4 -/- BM displayed a lack of organized vasculature, increased sinusoidal cavity size and a heightened expression of GPIbα + dysplastic and clustered MKs, particularly near the sinusoidal cavity. The findings emphasize the crucial role of β4galt1-dependent glycosylation in platelet production and distribution, fundamental for blood clotting and wound healing, during BM reconstitution after injury. Furthermore, the identification of the sinusoidal cavity as a reservoir of ECM and platelets, continuously remodeled during BM recovery, adds unexpected insights to our understanding of the BM reconstitution processes.

Prolonged oral antimicrobial administration prevents doxorubicin-induced loss of active intestinal stem cells

ABSTRACT Acute intestinal mucositis is a common off-target effect of chemotherapy, leading to co-morbidities such as vomiting, diarrhea, sepsis, and death. We previously demonstrated that the presence of enteric bacteria modulates the extent of jejunal epithelial damage induced by doxorubicin (DXR) in mice. Despite conventional thinking of the crypt as a sterile environment, recent evidence suggests that bacterial signaling influences aISC function. In this study, we labeled aISCs using transgenic Lgr5-driven fluorescence or with immunostaining for OLFM4. We examined the effect of DXR in both germ free (GF) mice and mice depleted of microbiota using an established antimicrobial treatment protocol (AMBx). We found differences in DXR-induced loss of aISCs between GF mice and mice treated with AMBx. aISCs were decreased after DXR in GF mice, whereas AMBx mice retained aISC expression after DXR. Neither group of mice exhibited an inflammatory response to DXR, suggesting the difference in aISC retention was not due to differences in local tissue inflammation. Therefore, we suspected that there was a protective microbial signal present in the AMBx mice that was not present in the GF mice. 16S rRNA sequencing of jejunal luminal contents demonstrated that AMBx altered the fecal and jejunal microbiota. In the jejunal contents, AMBx mice had increased abundance of Ureaplasma and Burkholderia. These results suggest pro-survival signaling from microbiota in AMBx-treated mice to the aISCs, and that this signaling maintains aISCs in the face of chemotherapeutic injury. Manipulation of the enteric microbiota presents a therapeutic target for reducing the severity of chemotherapy-associated mucositis.

Immune composition and its association with hematologic recovery after chemotherapeutic injury in acute myeloid leukemia

Chemotherapy-induced bone marrow (BM) injury is a significant cause of morbidity and mortality in acute myeloid leukemia (AML). Time to hematologic recovery after standard ("7+3") myeloablative chemotherapy can vary considerably among patients, but the factors that drive or predict BM recovery remain incompletely understood. Here, we assessed the composition of innate and adaptive immune subsets in the regenerating BM (day 17) after induction chemotherapy and related it to hematologic recovery in AML. T cells, and in particular the CD4 central memory (CD4CM) T-cell subset, were significantly enriched in the BM after chemotherapy, suggesting the relative chemoresistance of cells providing long-term memory for systemic pathogens. In contrast, B cells and other hematopoietic subsets were depleted. Higher frequencies of the CD4CM T-cell subset were associated with delayed hematopoietic recovery, whereas a high frequency of natural killer (NK) cells was related to faster recovery of neutrophil counts. The NK/CD4CM ratio in the BM after chemotherapy was significantly associated with the time to subsequent neutrophil recovery (Spearman's ρ=-0.723, p < 0.001, false discovery rate <0.01). The data provide novel insights into adaptive immune cell recovery after injury and identify the NK/CD4CM index as a putative predictor of hematopoietic recovery in AML.

Protein S-nitrosylation regulates proteostasis and viability of hematopoietic stem cell during regeneration

SUMMARYHematopoietic stem cells (HSCs) regenerate blood cells upon hematopoietic injuries. During homeostasis, HSCs are maintained in a low reactive oxygen species (ROS) state to prevent exhaustion. However, the role of nitric oxide (NO) in controlling HSC regeneration is still unclear. Here, we find increased NO during HSC regeneration with an accumulation of protein aggregation. S-nitrosoglutathione reductase (GSNOR)-deleted HSCs exhibit a reduced reconstitution capacity and loss of self-renewal after chemotherapeutic injury, which is resolved by inhibition of NO synthesis. Deletion of GSNOR enhances protein S-nitrosylation, resulting in an accumulation of protein aggregation and activation of unfolded protein response (UPR). Treatment of taurocholic acid (TCA), a chemical chaperone, rescues the regeneration defect of Gsnor−/− HSCs after 5-fluorouracil (5-FU) treatment. Deletion of C/EBP homologous protein (Chop) restores the reconstitution capacity of Gsnor−/− HSCs. These findings establish a link between S-nitrosylation and protein aggregation in HSC in the context of blood regeneration.

Lower proximal cup and outer root sheath cells regenerate hair bulbs during anagen hair follicle repair after chemotherapeutic injury.

The cell dynamics and cell origin for anagen hair follicle (HF) repair following chemotherapeutic injury are unclear. We first mapped the HF response to cyclophosphamide (CYP) at natural anagen VI in mice. We found that 30-60mg/kg of CYP leads to dose-dependent HF dystrophy that was spontaneously repaired with anagen resumption, while 120mg/kg of CYP prematurely induced catagen/telogen entry. To explore how anagen HF repair is achieved in the dystrophic anagen pathway, we analysed the cell dynamics at 30mg/kg of CYP. Hair bulbs first shrunk due to matrix cell apoptosis associated with DNA double-strand breaks. DNA damage was repaired, and ordered hair bulb structures were restored within 96hours. Bulge stem cells did not undergo apoptosis nor proliferation. K5+ basal lower proximal cup cells and outer root sheath cells quickly replenished the cells in the germinative zone and regenerated the concentric layered structures of the lower HF segment. Therefore, anagen HFs are able to summon extra-bulge progenitor cells in close proximity to the damaged matrix for quick repair after CYP injury.

Effects of chronic unpredictable mild stress on ovarian reserve in female rats: Feasibility analysis of a rat model of premature ovarian failure.

Premature ovarian failure (POF) results from a number of disorders. The POF model is primarily based on chemotherapeutic injury, and hence is not suitable for assessing the effects of chronic stress on ovarian function. Therefore, improved animal models are required to analyze the effects of chronic stress on ovarian reserve. The feasibility of the chronic unpredictable mild stress (CUMS) method for establishing a model of POF was examined. The depressive behavior exhibited by rats was evaluated with the open field and sucrose preference tests. Vaginal smears were obtained for assessment of the estrous cycle. The ovarian reserve of the animals was evaluated using the estrous cycle, ovarian histology and serum levels of gonadotropin releasing hormone (GnRH), follicle‑stimulating hormone (FSH), estradiol (E2), and anti‑Müllerian hormone (AMH). Compared with the control group, body weight, time spent in the center, horizontal movement, vertical frequency, consumption of sucrose, sucrose preference, number of small follicles from the rats, and serum E2, AMH and GnRH levels were significantly decreased in the CUMS group (all P<0.05). However, the estrous cycle was prolonged significantly (P<0.05) and serum FSH levels were increased significantly (P<0.01). These results suggested that the CUMS model rats exhibited depression‑like behaviors. CUMS may induce psychological stress and decrease ovarian reserve in female rats. Thus, the CUMS model may be used to assess the effects of chronic stress on female reproductive function.

Characterization of Endothelial Cells Associated with Hematopoietic Niche Formation in Humans Identifies IL-33 As an Anabolic Factor

Bone marrow formation requires an orchestrated interplay between osteogenesis, angiogenesis, and hematopoiesis that is thought to be mediated by endothelial cells. The nature of the endothelial cells and the molecular mechanisms underlying these events remain unclear in humans. Here, we identify a subset of endoglin-expressing endothelial cells enriched in human bone marrow during fetal ontogeny and upon regeneration after chemotherapeutic injury. Comprehensive transcriptional characterization by massive parallel RNA sequencing of these cells reveals a phenotypic and molecular similarity to murine type H endothelium and activation of angiocrine factors implicated in hematopoiesis, osteogenesis, and angiogenesis. Interleukin-33 (IL-33) was significantly overexpressed in these endothelial cells and promoted the expansion of distinct subsets of hematopoietic precursor cells, endothelial cells, as well as osteogenic differentiation. The identification and molecular characterization of these human regeneration-associated endothelial cells is thus anticipated to instruct the discovery of angiocrine factors driving bone marrow formation and recovery after injury.

Bone Marrow Myeloid Cells Regulate Myeloid-Biased Hematopoietic Stem Cells via a Histamine-Dependent Feedback Loop

Myeloid-biased hematopoietic stem cells (MB-HSCs) play critical roles in recovery from injury, butlittle is known about how they are regulated within the bone marrow niche. Here we describe an auto-/paracrine physiologic circuit that controls quiescence of MB-HSCs and hematopoietic progenitors marked by histidine decarboxylase (Hdc). Committed Hdc+ myeloid cells lie in close anatomical proximity to MB-HSCs and produce histamine, which activates the H2 receptor on MB-HSCs to promote their quiescence and self-renewal. Depleting histamine-producing cells enforces cell cycle entry, induces loss of serial transplant capacity, and sensitizes animals to chemotherapeutic injury. Increasing demand for myeloid cells via lipopolysaccharide (LPS) treatment specifically recruits MB-HSCs and progenitors into the cell cycle; cycling MB-HSCs fail to revert into quiescence in the absence of histamine feedback, leading to their depletion, while an H2 agonist protects MB-HSCs from depletion after sepsis. Thus, histamine couples lineage-specific physiological demands to intrinsically primed MB-HSCs to enforce homeostasis.

Angelica sinensis Polysaccharides Ameliorate Stress-Induced Premature Senescence of Hematopoietic Cell via Protecting Bone Marrow Stromal Cells from Oxidative Injuries Caused by 5-Fluorouracil.

Myelosuppression is the most common complication of chemotherapy. Decline of self-renewal capacity and stress-induced premature senescence (SIPS) of hematopoietic stem cells (HSCs) induced by chemotherapeutic agents may be the cause of long-term myelosuppression after chemotherapy. Whether the mechanism of SIPS of hematopoietic cells relates to chemotherapeutic injury occurred in hematopoietic microenvironment (HM) is still not well elucidated. This study explored the protective effect of Angelica sinensis polysaccharide (ASP), an acetone extract polysaccharide found as the major effective ingredients of a traditional Chinese medicinal herb named Chinese Angelica (Dong Quai), on oxidative damage of homo sapiens bone marrow/stroma cell line (HS-5) caused by 5-fluorouracil (5-FU), and the effect of ASP relieving oxidative stress in HM on SIPS of hematopoietic cells. Tumor-suppressive doses of 5-FU inhibited the growth of HS-5 in a dose-dependent and time-dependent manner. 5-FU induced HS-5 apoptosis and also accumulated cellular hallmarks of senescence including cell cycle arrest and typical senescence-associated β-galactosidase positive staining. The intracellular reactive oxygen species (ROS) was increased in 5-FU treated HS-5 cells and coinstantaneous with attenuated antioxidant capacity marked by superoxide dismutase and glutathione peroxidase. Oxidative stress initiated DNA damage indicated by increased γH2AX and 8-OHdG. Oxidative damage of HS-5 cells resulted in declined hematopoietic stimulating factors including stem cell factor (SCF), stromal cell-derived factor (SDF), and granulocyte-macrophage colony-stimulating factor (GM-CSF), however, elevated inflammatory chemokines such as RANTES. In addition, gap junction channel protein expression and mediated intercellular communications were attenuated after 5-FU treatment. Significantly, co-culture on 5-FU treated HS-5 feeder layer resulted in less quantity of human umbilical cord blood-derived hematopoietic cells and CD34+ hematopoietic stem/progenitor cells (HSPCs), and SIPS of hematopoietic cells. However, it is noteworthy that ASP ameliorated SIPS of hematopoietic cells by the mechanism of protecting bone marrow stromal cells from chemotherapeutic injury via mitigating oxidative damage of stromal cells and improving their hematopoietic function. This study provides a new strategy to alleviate the complication of conventional cancer therapy using chemotherapeutic agents.

The Squid Ink Polysaccharides Protect Tight Junctions and Adherens Junctions from Chemotherapeutic Injury in the Small Intestinal Epithelium of Mice

Gastrointestinal mucositis and infection by chemotherapy treatment are associated with alterations in the functioning of the intestinal barrier, due to the potential damage induced by anticancer drugs on the epithelial tight junctions and adheren junction. We aimed to study the protective effect of dietary polysaccharides on chemotherapy-induced injury in the epithelial cells. In the current study, using mice that were intraperitoneally injected with 50 mg/kg cyclophosphamide for 2 days, we reveal that polysaccharides from the ink of Ommastrephes bartrami (OBP) enhanced the mRNA and protein expression levels of Occludin, zonulae occluden (ZO)-1, and E-cadherin. Immunohistochemistry staining of ZO-1 and E-cadherin confirmed the increase in the mRNA and protein levels. OBP also remarkably enhanced the mRNA expression of other tight junction proteins, ZO-2, ZO-3, claudin-2, and cingulin. Our results may have important implications in host defense, especially the immunopotentiation function of OBP on the cyclophosphamide-induced epithelial cell injury, as well as intestinal disorders involving inflammation and infection.

The dietary polysaccharide from Ommastrephes bartrami prevents chemotherapeutic mucositis by promoting the gene expression of antimicrobial peptides in Paneth cells

Gastrointestinal mucositis (GIM), induced by chemotherapy, is associated with alterations in the function of the intestinal barrier due to the potential damage induced by anti-cancer drugs on the epithelial cells. The Paneth cells, an important epithelial lineage in the intestine, contribute to the innate immunity by releasing antimicrobial proteins onto the mucosal surfaces. Polysaccharide, as a widely used immunomodulatory nutrient, was reported to have an effect of modulating the intestinal immune system. However, the effect of dietary polysaccharides in protecting the Paneth cells from injuries induced by chemotherapy was poorly investigated. In the current study, with a mouse model of cyclophosphamide (Cy)-induced gastrointestinal injury, the polysaccharide extracted from the ink of Ommastrephes bartrami (OBP) were found to protect the mouse Paneth cells from chemotherapeutic injury. Ommastrephes bartrami polysaccharide (OBP) treatment could enhance the mRNA expression of antimicrobial proteins in the Paneth cells, such as lysozyme, Ang4, Defa-5, and sPLA2. The increased production of the microbicidal proteins in the Paneth cells was dependent on the relatively highly developed endoplasmic reticulum (ER) structure but was not dependent on the increase in the quantity of the ER. The Ommastrephes bartrami polysaccharide (OBP) activated the IRE-1 mediated XBP-1s pathway to cope with antimicrobial protein secretion. Our results may have important implications with regard to the role of OBP in chemotherapy-induced Paneth cell injury in intestinal disorders involving inflammation and infection.

Dietary squid ink polysaccharide could enhance SIgA secretion in chemotherapeutic mice.

Secretory immunoglobulin A (SIgA) is a non-inflammatory antibody that shields internal body surfaces, such as in the intestine to neutralize pathogens in the lumen of the intestine. As chemotherapy seriously damages the mucosal immune system, we herein demonstrated that polysaccharide from the squid ink of Ommastrephes bartrami (OBP) activated intestinal SIgA secretion to prevent chemotherapeutic injury. Using a mouse model of chemotherapy induced intestinal injury by intraperitoneal injection of 50 mg kg(-1) cyclophosphamide, our results showed an enhanced SIgA concentration in intestinal mucosa by OBP administration and the higher production of SIgA relied on the greater expression of IgA, J chain and pIgR. Furthermore, the higher expressions of IL-6, IL-10 and TNF-α increased by OBP treatment contributed to enhanced IgA and J chain synthesis in IgA(+) plasma cells, and pIgR expression in epithelial cells. It also triggered a prompt immunoglobulin secretory pathway confirmed by enhanced UPR (unfolded protein response) effectors XBP-1s and Bip expression. Our results have important implications for the mucosal immunity enhancement effects of OBP as a functional food component for chemotherapeutic patients.

Left Superior Hepatectomy and Segment 6 Resection for Colorectal Cancer Metastasis Invading the Left Hepatic Vein: An Actual Parenchyma Preserving Technique

Introduction: Hepatic metastasis is the most common form of distant spread of colorectal cancer (CRC) with about 50% occurrence rate. Liver resection (LR) with R0 margins is the only curative treatment and is believed to have improved the long-term out-come of these patients. Because of a chemotherapeutic injury to the liver, preservation of as much parenchymal volume as possible to minimize the risk of liver failure is the most important issue in these group of patients.Our present report describes a parenchyma preserving technique with left superior hepatectomy and segment 6 resection in a case. Case Report: A 64- year- old woman presented to our instution with a colorectal liver metastasis. PET-CT scan showed solitary liver lesions in segment 2-4a and 6. A left superior hepatectomy (segment 2 and 4a) and segment 6 resection was performed with glissonian approach and clemp-crush technique. Left hepatic vein was ligated without blocking the venous and biliary drainage of segment 4b and segment 3. Pathological examination of the specimen showed tumor-free margins (R0 resection). Discussion: Developments in imaging modalities provide an improved visualization of hepatic segmental anatomy and also provide volumetric calculation on the liver. This allows a successful planning for segmental liver resections with a minimum risk of postoperative liver failure. Factors that were considered contraindications for the surgery, such as number of metastases, tumor size, synchronous metastases and the presence of extrahepatic disease, must be evaluated as prognostic factors and must not prevent these patients opportunity of being treated. The main consideration is to achieve a complete R0 resection. A 1cm-R0 surgical margin width has been considered to avoid local intrahepatic recurrence and optimize long-term survival after hepatic resection for colorectal cancer metastases but tumor biology is a more important predictor for intrahepatic recurrence rather than milimetres. Conclusion: Preservation of as much parenchymal volume as possible in order to minimize the risk of liver failure is required in liver resections with chemotherapeutic liver injury. Drainage of the remaining liver segments into the retrohepatic vena cava via the retrohepatic veins and communicating veins between adjacent hepatic veins may allow adequate liver outflow and remaining functional liver parenchyma in selected cases with hepatic vein invasion.

An In Vivo Functional Screen Identifies miRNA-150 As a Regulator of Hematopoietic Regeneration Post Chemotherapeutic Injury

Abstract 2333.MicroRNAs are important regulators of many hematopoietic processes, yet little is known with regard to the role of microRNAs in controlling normal hematopoietic regeneration. The most common methodology for in vivo microRNA studies follows a hypothesis-driven candidate approach. Here, we report the establishment of an unbiased, in vivo, microRNA gain-of-function screen, and the identification of miR-150 as a negative regulator of hematopoietic recovery post chemotherapeutic challenge.Specifically, a retroviral-library consisting of 135 hematopoietic-expressed microRNAs was generated, with each expression construct containing a barcode sequence that can be specifically recognized using a novel bead-based platform. Hematopoietic-stem-and-progenitor-cell (HSPC)-enriched wild-type bone marrow was transduced with this library and transplanted into lethally-irradiated recipients. Analysis of peripheral blood samples from each recipient up to 11 weeks post transplantation revealed that 87% of the library barcodes are reliably detected. To identify microRNAs that regulate hematopoietic regeneration after chemotherapy-induced injury, we measured the change in barcode abundance for specific microRNA constructs after 5-fluorouracil (5-FU) challenge. Notably, a small number of barcodes were consistently depleted in multiple recipient mice after treatment. Among the top hits was the miR-150-associated barcode, which was selected for further experimentation. Indeed, overexpression of miR-150 in a competitive environment resulted in significantly lower recovery rates for peripheral myeloid and platelet populations after 5-FU treatment, whereas the effects on B- and T-cells were milder. Furthermore, full recovery of these cell populations did not occur until ∼12 weeks after treatment, suggesting the involvement of HSPCs and/or common lineage progenitors. Conversely, knocking out miR-150 led to an opposite phenotype, with platelets and myeloid cells displaying faster recovery in both competitive and non-competitive settings. Interestingly, we could not observe the described effects of miR-150 in bone marrow primary cell cultures, suggesting that such effects cannot be recapitulated in vitro.Overall, these data indicate that miR-150 is a novel regulator of hematopoietic recovery after chemotherapeutic-induced injury, and highlight the important role of microRNAs in the intrinsic wiring of the hematopoietic regeneration program. Our experiments also demonstrate the feasibility and power of functional in vivo screens for studying normal hematopoietic functions, which can become an important tool in the hematology field. Disclosures:No relevant conflicts of interest to declare.

In vivo and in vitro effect of peptide HP-6 derived from donkey serum albumin on hematopoietic system

By bioinformatics method, the effect in hematopoietic system of bioactive peptide HP-6, which was obtained from donkey serum albumin and is one of the major protein components from donkey-hide gelatin, was investigated. Human bone marrow nucleated cells (hBMNCs) and murine bone marrow stromal cells (mBMSCs) were separated and cultured with different concentration of peptide HP-6 (0.000 15, 0.001 5, 0.015, 0.15, 1.5 micromol x L(-1)). The effect on promoting proliferation of cells related to hematopoiesis in bone morrow was detected and the ultrastructure of cells after treated by HP-6 was observed through transmission electron microscope. Hemorrhage anemia mouse model and anemia mouse model induced by cyclophosphamide were established, and randomly divided into peptide HP-6 groups which were administered respectively with different doses (1, 0.1, 0.01 mg x kg(-1)) by gavage, and control group which was administered with PBS by gavage. Peripheral blood components of all mice and bone morrow cells (BMC) number of mice induced by cyclophosphamide were evaluated. Peptide HP-6 could concentration-related promote the proliferation of hBMNCs and mBMSCs, hBMNCs got the highest reproduction rate of 152.11% and mBMSCs also got 63.52% with the concentration of 0.15 micromol x L(-1), then the reproduction rate decreased while the concentration kept increasing. The transmission electron microscope showed that ultrastructure of cells was normal after treated by HP-6.1 mg x kg(-1) peptide HP-6 significantly increased peripheral platelet and protected mouse morrow injured by cyclophoshamide. 0.1 mg x kg(-1) peptide HP-6 significantly increased peripheral platelet with relative growth rate of 77.65%, increased peripheral white blood cells count and peripheral red blood cells count, also could protect mouse peripheral blood after treated by chemotherapeutics. Peptide HP-6 could promote the proliferation of cells related to hematopoietic system, enhance mouse hemopoiesis function and the resistance to chemotherapeutic injury.

Clinical scoring systems for predicting outcome after surgery for colorectal liver metastases: towards a better multidisciplinary approach

Clinical scoring systems for predicting outcome after surgery for colorectal liver metastases: towards a better multidisciplinary approach

Cranial mesohepatectomy: a challenging parenchyma-preserving operation for colorectal liver metastases

Dear Editor: The treatment of colorectal liver metastases has evolved over time. Improvements in chemotherapeutic regimens coupled with a multidisciplinary approach have resulted in a greater number of patients undergoing liver resections and major surgical challenges. Chemotherapeutic injury to the liver mandates preservation of as much parenchymal volume as possible in order to minimize the risk of liver failure. Furthermore, the potential for second and third resections for recurrent disease mandates the maintenance as much as possible of the intrahepatic anatomical relationships. Our present report describes a cranial mesohepatectomy in a case considered to be inoperable by others. A 57-year-old woman presented to our institution with a colorectal liver metastasis. Approximately 1 year ago, she had undergone a low anterior resection for a nodal negative rectal adenocarcinoma (pT3N0V0L0R0G2). Adjuvant radiochemotherapy according to the MAYO protocol had been initiated but side effects led to its discontinuation after the first cycle. A computed tomography (CT) scan at the time of presentation showed a 19 cm in diameter central solitary liver lesion infiltrating the right and middle hepatic veins. A right trisectionectomy (resection of segments 4– 8) was considered but not pursued due to the high risk of postoperative liver insufficiency (calculated remnant liver volume of 215 ml). Careful study of CT images showed a right inferior hepatic vein draining part of the lateral sector of the right liver lobe. Virtual postresection reconstruction with computer-assisted three-dimensional CT showed adequate drainage of segments 5 and 6 and partial drainage of segment 4b (379 ml) via the right inferior hepatic vein. A cranial mesohepatectomy (segments 4a, 7, and 8) was undertaken. The lesion was resected en bloc with the right and middle hepatic veins. Pathological examination of the specimen showed tumor-free margins (R0 resection). The patient tolerated the operation well, with no signs of liver insufficiency. She was discharged on postoperative day 10. Surgical resection of colorectal liver metastases requires precise anatomical study of the individual intrahepatic anatomy. Recent developments in computer-assisted imaging provide improved visualization of the hepatic vascular and segmental anatomy and allow for volumetric calculation. Image-based computer assistance and three-dimensional reconstruction are particularly useful for planning challenging liver resections, such as cranial mesohepatectomy, minimizing the risk of postoperative liver failure. Int J Colorectal Dis (2009) 24:243 DOI 10.1007/s00384-008-0610-1