Impact of malnutrition on cancer recurrence, colorectal cancer-specific death, and non-colorectal cancer-related death in patients with colorectal cancer who underwent curative surgery.

AIM: This study aimed to examine how malnutrition, as reflected by the Geriatric Nutritional Risk Index (GNRI), is associated with colorectal cancer (CRC) recurrence and cause of death. METHODS: Consecutive stage I-III CRC patients (n = 601) were divided into two groups using GNRI 98 as the cutoff. The relationship of GNRI with overall survival (OS) and recurrence-free survival (RFS) was evaluated, followed by competing risk analysis to determine prognostic factors of non-CRC-related death, and hazard function analysis to examine changes in the risk of recurrence and death. RESULTS: Median body mass index was lower in the low GNRI group than in the high GNRI group (19.8 vs. 23.5; p < 0.001). After adjusting for known prognostic factors, a low GNRI was independently associated with reduced OS/RFS, and was a significant predictor of non-CRC-related death. The risk of recurrence was higher and peaked earlier in the low GNRI group than in the high GNRI group, although after 3 years, both groups had a similar risk. Meanwhile, the low GNRI group had a higher risk of non-CRC-related death over the course of 5 years. CONCLUSION: It is important to consider preoperative nutritional status along with the cancer stage when developing strategies to improve outcomes for CRC patients.

Level of Inferior Mesenteric Artery Ligation in Sigmoid Colon and Rectal Cancer Surgery: Analysis of Apical Lymph Node Metastasis and Recurrence.

INTRODUCTION: Whether high or low ligation of the inferior mesenteric artery (IMA) is optimal for treating sigmoid colon and rectal cancers is controversial. The present study aimed to compare outcomes of high and low ligation of the IMA and determine the adequate extent of IMA lymph node dissection. METHODS: Subjects were 455 consecutive stage I-III colorectal cancer patients who underwent curative surgery between 2011 and 2019. We assessed the association between the level of IMA ligation and overall survival and recurrence-free survival (RFS) by propensity score matching analysis. Clinicopathological features of IMA lymph node metastasis and recurrence patterns were analyzed. RESULTS: After propensity score matching, the low ligation group had a significantly worse prognosis than that of the high ligation group for RFS (p = 0.039). Positive IMA lymph nodes were associated with pathological T3 or T4 stage and N2 stage. IMA lymph node recurrences in the high ligation group occurred at the superior left side of the IMA root. In contrast, all recurrences in the low ligation group occurred at the left colic artery bifurcation. CONCLUSION: High ligation of IMA is oncologically safe. However, even with high ligation, care must be taken to ensure adequate lymph node dissection.

Novel and efficient method for culturing patient-derived gastric cancer stem cells.

Experimental techniques for patient-derived cancer stem-cell organoids/spheroids can be powerful diagnostic tools for personalized chemotherapy. However, establishing their cultures from gastric cancer remains challenging due to low culture efficiency and cumbersome methods. To propagate gastric cancer cells as highly proliferative stem-cell spheroids in vitro, we initially used a similar method to that for colorectal cancer stem cells, which, unfortunately, resulted in a low success rate (25%, 18 of 71 cases). We scrutinized the protocol and found that the unsuccessful cases were largely caused by the paucity of cancer stem cells in the sampled tissues as well as insufficient culture media. To overcome these obstacles, we extensively revised our sample collection protocol and culture conditions. We then investigated the following second cohort and, consequently, achieved a significantly higher success rate (88%, 29 of 33 cases). One of the key improvements included new sampling procedures for tumor tissues from wider and deeper areas of gastric cancer specimens, which allowed securing cancer stem cells more reproducibly. Additionally, we embedded tumor epithelial pieces separately in both Matrigel and collagen type-I as their preference to the extracellular matrix was different depending on the tumors. We also added a low concentration of Wnt ligands to the culture, which helped the growth of occasional Wnt-responsive gastric cancer stem-cell spheroids without allowing proliferation of the normal gastric epithelial stem cells. This newly improved spheroid culture method may facilitate further studies, including personalized drug-sensitivity tests prior to drug therapy.

Impact of severe postoperative inflammatory response on recurrence after curative resection of colorectal cancer.

PURPOSE: The relationship between recurrence and systemic inflammation in the early postoperative period of colorectal cancer (CRC) is unclear, and no study has examined changes in recurrence risk resulting from postoperative inflammation. This study aimed to investigate the prognostic impact of severe postoperative inflammatory response after curative resection of CRC. METHODS: Subjects were 422 consecutive CRC patients who underwent curative surgery between 2012 and 2016, with a follow-up period of 5 years after surgery. The prognostic impacts of high and low postoperative C-reactive protein (CRP) levels were examined, and temporal changes in recurrence risk were examined using hazard function analysis. RESULTS: Patients were divided into high (n = 68) and low (n = 354) CRP groups using CRP 12.5 mg/dL as the cut-off based on receiver-operating characteristic (ROC) curve analysis. Multivariable analysis revealed that high CRP was a significant predictor of recurrence (hazard ratio: 2.21; p < 0.001). According to the hazard function of recurrence risk, the hazard rate of the low CRP group peaked at 8.4 months postoperatively (peak hazard rate: 0.0073/month) and decreased thereafter, while that of the high CRP group peaked at 5.6 months (0.0142/month) and was persistently higher than that of the low CRP group for the study duration. CONCLUSIONS: A severe inflammatory response in the early postoperative period of CRC increased the recurrence risk, and the recurrence hazard of patients with high CRP levels was earlier and persistently higher than that of patients with low CRP levels. CRC patients with high CRP levels may require more frequent follow-up.

Association of nutritional risk and systemic inflammation with survival in patients with colorectal cancer who underwent curative surgery.

BACKGROUND & AIMS: Malnutrition and inflammation adversely affect the prognosis of patients with cancer. The Geriatric Nutritional Risk Index (GNRI) and systemic inflammatory markers, including neutrophil-to-lymphocyte ratio (NLR), lymphocyte-to-monocyte ratio (LMR), lymphocyte-to-C reactive protein ratio (LCR), and C-reactive protein-to-albumin ratio (CAR), predict survival in colorectal cancer (CRC) patients. The present study aimed to examine the association of these two factors with CRC survival. METHODS: Subjects were 433 consecutive CRC patients who underwent curative surgery between 2013 and 2018. Patients were stratified by nutritional status, and relationships between overall survival (OS) and systemic inflammation were evaluated. The prognostic impact of combinations of the GNRI and inflammatory markers was assessed. Multivariable analyses were also performed. RESULTS: All assessed biomarkers predicted OS in univariable analysis (GNRI:P < 0.001, NLR:P = 0.048, LMR:P = 0.001, LCR:P = 0.010, CAR: P = 0.039). Stratified analysis showed that each inflammatory marker had a prognostic impact on OS in the low GNRI group (NLR:P = 0.028, LMR:P = 0.003, LCR:P = 0.05, CAR:P = 0.009). In contrast, inflammatory markers had no prognostic impact on OS in the high GNRI group. The combination of malnutrition and systemic inflammation had a high prognostic value (all P < 0.016). The multivariable analysis revealed that a low GNRI (hazard ratio: 2.58-2.89) was independently associated with reduced survival, whereas none of the inflammatory markers independently predicted poor prognosis. CONCLUSIONS: The GNRI is a useful prognostic biomarker for CRC patients, whereas systemic inflammatory markers can only serve as prognostic factors when patient nutritional status is taken into account. The combination of malnutrition and systemic inflammation may enhance the accuracy of prognostic prediction.

Appropriate traction can help identify the optimal dissectable layer for infrapyloric lymph node dissection in laparoscopic gastrectomy.

INTRODUCTION: Appropriate dissection of the infrapyloric lymph nodes (no. 6 LNs) is important in gastric cancer surgery. In laparoscopic surgery, dissection of the no. 6 LNs along the inner dissectable layer from the left side of patient has been reported. However, it is difficult for surgeons to provide appropriate traction with their left hand from the left side. To resolve this difficulty, we dissected the no. 6 LNs from the patient's right side to identify the optimal layer. We then evaluated the oncologic reliability of the layer and the safety of this procedure. METHODS: From the patient's right side, the surgeon used their left hand to provide appropriate traction when pulling the adipose tissue, including the no. 6 LNs. This exposed the optimal layer between the adipose tissue and the pancreas. To assess this maneuver, the surgical outcomes of patients who underwent laparoscopic distal gastrectomy from April 2011 to March 2013 were retrospectively analyzed. The surgical outcomes included the number of the no. 6 LNs resected, time to dissect the no. 6 LNs, incidence of pancreatic complications, and recurrence in the no. 6 LNs. RESULTS: There were 112 patients identified. The median number of the no. 6 LNs resected was five. The median time to dissect the no. 6 LNs was 14 minutes. Four patients developed pancreatic fistula, and another four patients developed intra-abdominal abscess. There was no recurrence in the no. 6 LNs. CONCLUSION: The optimal layer was oncologically reliable, and these procedures were safe.

Chemosensitivity of Patient-Derived Cancer Stem Cells Identifies Colorectal Cancer Patients with Potential Benefit from FGFR Inhibitor Therapy.

Some colorectal cancer patients harboring FGFR (fibroblast growth factor receptor) genetic alterations, such as copy number gain, mutation, and/or mRNA overexpression, were selected for enrollment in several recent clinical trials of FGFR inhibitor, because these genetic alterations were preclinically reported to be associated with FGFR inhibitor sensitivity as well as poor prognosis, invasiveness, and/or metastatic potential. However, few enrolled patients were responsive to FGFR inhibitors. Thus, practical strategies are eagerly awaited that can stratify patients for the subset that potentially responds to FGFR inhibitor chemotherapy. In the present study, we evaluated the sensitivity to FGFR inhibitor erdafitinib on 25 patient-derived tumor-initiating cell (TIC) spheroid lines carrying wild-type RAS and RAF genes, both in vitro and in vivo. Then, we assessed possible correlations between the sensitivity and the genetic/genomic data of the spheroid lines tested. Upon their exposure to erdafitinib, seven lines (7/25, 28%) responded significantly. Normal colonic epithelial stem cells were unaffected by the inhibitors. Moreover, the combination of erdafitinib with EGFR inhibitor erlotinib showed stronger growth inhibition than either drug alone, as efficacy was observed in 21 lines (84%) including 14 (56%) that were insensitive to erdafitinib alone. The in vitro erdafitinib response was accurately reflected on mouse xenografts of TIC spheroid lines. However, we found little correlation between their genetic/genomic alterations of TIC spheroids and the sensitivity to the FGFR inhibitor. Accordingly, we propose that direct testing of the patient-derived spheroids in vitro is one of the most reliable personalized methods in FGFR-inhibitor therapy of colorectal cancer patients.

A Chemosensitivity Study of Colorectal Cancer Using Xenografts of Patient-Derived Tumor-Initiating Cells.

Current genomic and gene expression analyses provide versatile tools to improve cancer chemotherapy. However, it is still difficult to predict whether each patient responds to a particular regimen or not. To predict chemosensitivity in each patient with colorectal cancer, we developed an evaluation method using the primary tumor-initiating cells (TIC, aka cancer stem cells) xenografted in nude mice subcutaneously (patient-derived spheroid xenografts; PDSX). Simultaneously, we also prepared the conventional patient-derived xenografts (PDX) from the same patients' tumors and compared the dosing results with those of PDSXs. We further compared the chemosensitivities of PDSXs with those of 7 patients who had been given regimens such as FOLFOX and FOLFIRI to treat their metastatic lesions. As per the results, the PDSX method provided much more precise and predictable tumor growth with less variance than conventional PDX, although both retained the epithelial characteristics of the primary tumors. Likewise, drug-dosing tests showed essentially the same results in PDXs and PDSXs, with stronger statistical power in PDSXs. Notably, the cancer chemosensitivity in each patient was precisely reflected in that of the PDSX mice along the clinical course until the resistance emerged at the terminal stage. This "paraclinical" xenograft trials using PDSXs may help selection of chemotherapy regimens efficacious for each patient, and, more importantly, avoiding inefficient ones by which the patient can lose precious time and QOL. Furthermore, the PDSX method may be employed for evaluations of off-label uses of cancer chemotherapeutics and compassionate uses of yet-unapproved new drugs in personalized therapies. Mol Cancer Ther; 17(10); 2187-96. ©2018 AACR.

An improved method for culturing patient-derived colorectal cancer spheroids.

Recent advances allowed culturing and examination of patient-derived colorectal cancer (PD-CRC) cells as organoids or spheroids. To be applied to practical personalized medicine, however, current methods still need to be strengthened for higher efficiency. Here we report an improved method to propagate PD-CRC tumor initiating cells (TICs) in spheroid culture. We established > 100 cancer spheroid lines derived from independent colorectal cancer patients employing a serum-containing medium with additional inhibitors, Y27632 and SB431542. Because colorectal cancer spheroids showed wide-range growth rates depending on the patient tumors, we searched for supplementary factors that accelerated proliferation of slow-growing CRC-TIC spheroids. To this end, we introduced a convenient growth-monitoring method using a luciferase reporter. We found that epidermal growth factor (EGF) and/or basic fibroblast growth factor (bFGF) were critical for steady propagation of a subset of CRC-TIC spheroids carrying the wild-type RAS and RAF genes. We also identified 5'-(N-ethyl-carboxamido)-adenosine (NECA), an adenosine receptor agonist, as an essential supplement for another subset of spheroids. Based on these results, we propose to optimize culture conditions for CRC-TIC spheroids by adjusting to the respective tumor samples. Our method provides a versatile tool that can be applied to personalized chemotherapy evaluation in prospective clinical studies.

Accurate diagnosis of mismatch repair deficiency in colorectal cancer using high-quality DNA samples from cultured stem cells.

Mismatch repair (MMR)-deficient or microsatellite instability (MSI) colorectal cancer includes two subtypes; Lynch syndrome and sporadic MSI cancer, both of which generate multiple neoantigens due to unrepaired mutations. Although such patients respond very well to immune checkpoint therapy, their diagnosis can be confused by low quality DNA samples owing to formalin fixation and/or low cancer cell content. Here we prepared high-quality DNA samples from in vitro-cultured cancer spheroids that consisted of the pure cell population. We evaluated their diagnostic power by on-chip electrophoresis, mutational burden assessment, and direct sequencing. Because formalin-fixed paraffin-embedded (FFPE) tissues are widely used as the DNA source, we compared such samples with spheroid DNA. Additionally, we performed immunohistochemistry (IHC) for MMR proteins on spheroids as well as primary tumor sections. Of 111 cases of colorectal cancer patients, we found seven MSI-high cases in which all diagnostic results agreed on spheroid-based assays, whereas the results with the FFPE DNA were less reliable though analyzable. Importantly, there was an MSS case that appeared as MSI by IHC on primary tumor sections. Based on these results, we propose to employ cultured cancer spheroids as the source of both DNA and IHC specimens for more reliable clinical diagnosis.

Esophagogastric tube reconstruction with stapled pseudo-fornix in laparoscopic proximal gastrectomy: a novel technique proposed for Siewert type II tumors.

PURPOSE: The incidence of adenocarcinoma of the esophagogastric junction is increasing, but laparoscopic proximal gastrectomy is not widely accepted due to the absence of a standardized technique of reconstruction. This report describes a novel technique of esophagogastric tube reconstruction in laparoscopic proximal gastrectomy for Siewert type II tumors. METHODS: Laparoscopic proximal gastrectomy, sometimes with transhiatal distal esophagectomy, was performed. After a perigastric, suprapancreatic, and lower thoracic paraesophageal lymphadenectomy, a gastric tube of 35-mm width was prepared. An esophagogastric tube anastomosis with pseudo-fornix was made with a no-knife linear stapler to prevent postoperative reflux esophagitis. RESULTS: Fifteen patients with Siewert type II tumors underwent this operation. They included six patients with early-stage cancer, six at high risk for transhiatal total gastrectomy due to several comorbidities, and three who needed palliative tumor resection. The mean operation time was 315 min. One postoperative anastomotic leak was treated conservatively, and three anastomotic stenoses were resolved with endoscopic balloon dilatation. Postoperative 1-year follow-up endoscopy revealed four cases of reflux esophagitis that were well controlled by medication. CONCLUSIONS: This new technique of reconstruction was feasible. With the advantage of a gastric tube, a tension-free anastomosis was possible even for bulky tumors that needed lower esophagectomy. Although long-term follow-up and a larger number of patients are required to evaluate long-term functional outcomes and oncological adequacy, our procedure has the potential of becoming a treatment of choice for early-stage Siewert type II tumors and/or for some selected high-risk patients who need tumor resection.

[A case of intraductal mucinous neoplasms with the whole main pancreatic duct dilation treated via segment pancreatectomy].

The patient was a 77-year-old woman. She was diagnosed as intraducal papillary mucinous neoplasms (IPMN). She refused an operation for 3 years. After all, a nodule in the main pancreatic duct was pointed out, she agreed and was referred to us. Her past history showed pacemaker implantation for third-degree atrioventricular block, and no impaired glucose tolerance. Abdominal CT showed a dilated whole pancreatic duct and a multilocular cystic tumor. Endoscopic retrograde pancreatography showed a marked dilation of the main pancreatic duct. We diagnosed as main duct IPMN. Intraoperative US showed no nodule in pancreatic duct, and there was no suspicious lesion of invasive cancer. We performed segmental pancreatectomy between the left side of common bile duct and the pancreas tail. The tumor was resected with clear margins. Both cut-ends of the main pancreatic duct were anastomosed to a jejunal loop. The postoperative course was excellent. She was discharged on day 16. The glycemic control was good, she needed no treatment for diabetes. Total pancreatectomy has many problems such as insulin and pancreatic polypeptide deficiency, hypoglycemia, malabsorption, diarrhea and liver dysfunction. We avoided total pancreatectomy so that her quality of life was maintained. Still a careful follow -up is required.

[Multimodal treatment for MFH originating from the psoas muscle-a case report].

A 51-year-old male presented with lower abdominal mass and thigh and lumbar pain. CT scan showed a large retroperitoneal tumor compressing the lumbar vertebrae and IVC. Arterial infusion of mitomycin C, epirubicin and DSM was performed for unresectable tumor. The tumor was shrunken and the infusion repeated in 6 and 10 weeks later until right femoral palsy occurred. Radical resection with lumbar fascicles and psoas muscle and IORT (25 Gy) was performed. Local recurrence appeared before the sacrum in the next year. Arterial infusion and resection was done. Local recurrence invading the right common iliac artery was found five years later. The tumor and the artery were resected, followed by extra-anatomical ilio-iliac bypass. Local recurrences occurred in the 6th, 7th and 9th year. Radiation therapy was effective. But for the last recurrence, no radiation was applicable because of over dosage. As the most important thing in the treatment of MFH is to resect the tumor with curative margin at the initial operation, and the retroperitoneum and pelvic cavity are not good places to obtain good margins, multimodal treatment including irradiation should have been considered even preoperatively. Also consultation with orthopedic oncologists, radiologists, vascular surgeons and team practice are essential to get curative margins.

[Intramucosal colorectal carcinomas treated by surgical resection].

Stage 0 colorectal cancer was found only in the innermost lining of the colon and rectum. Treatments for an early stage colorectal cancer were available including endoscopic polypectomy, endoscopic mucosal resection (EMR) and trans-anal or -sacral local excision, laparoscopy-assisted colectomy and open colectomy. Our study indicated that endoscopic therapy for the early stage colorectal cancer was more advantageous than the conventional operative treatment. Although EMR should be applied for intramucosal carcinomas, 11 intramucosal carcinomas were treated by a surgical resection due to several limitations at our institution.

[A case of thrombocytopenia after microwave coagulation therapy for multiple metastatic liver tumors].

The patient was a 76-year-old woman. One year ago, she was diagnosed as ascending colon cancer with the multiple metastases. Chest and abdominal CT showed more than 50 metastases in the liver, lung metastasis and the lesser curvature lymph metastases. She was received chemotherapy (mFOLFOX6, 5-FU/LV regimen). After 10 months, CT showed disappearance of lung metastasis and a reduction of liver metastases. We diagnosed as resectable. She had undergone right hemicolectomy, dissection of the lesser curvature lymph nodes, partial hepatectomy (17 places) and microwave coagulonecrotic therapy for liver metastases (20 places). After the operation, the number of platelets decreased from 14 × 10(4) to 30 × 10(4)/mm3. Anticoagulant therapy was effective and the platelet count increased. Postoperative bleeding, infection and liver dysfunction were not confirmed. We assumed the cause of thrombocytopenia was local intravascular coagulation due to ablation of the liver.

[Successful treatment of multiple pulmonary and peritoneal recurrence of hepatocellular carcinoma with bronchial artery infusion therapy and PEIT followed by surgery--a case report].

A case of hepatocellular carcinoma, successfully treated with multimodal loco-regional treatments, is reported. An 80-year-old male presented with multiple pulmonary and peritoneal metastases 4 months after right heimihepatectomy for ruptured HCC. Bronchial artery infusion of mitomycin C induced pulmonary tumor regression and stabilization. Peritoneal tumor was treated by arterial infusion of SMANCS, followed by percutaneous injection of absolute ethanol, which ended in surgical removal in 28-postoperative month due to abscess formation. He had been well until right adrenal and left pulmonary metastases appeared. Resection of both metastases was carried out in 39-post hepatectomy month. Recurrent left pulmonary metastasis was treated with two sessions of bronchial artery infusion with no effect this time. Video-assisted partial resection of the left lung was performed in 54 post-hepatectomy month. But his AFP level kept rising. Eventually pulmonary metastasis recurred and tumor thrombus reached the left atrium 58 months after hepatectomy. He wanted no more treatment. He died of cerebral infarction caused by tumor thrombus. He enjoyed a good QOL for five years through multimodal loco-regional treatments.

[Case of bilateral subdural empyema complicating Campylobacter fetus subspecies fetus meningitis].

Campylobacter fetus subspecies fetus (C. fetus) is a relatively rare cause of meningitis, with only 3 cases of C. fetus subdural empyema reported to date. We experienced a case of bilateral subdural empyema complicating C. fetus meningitis. A 51-year-old man was found lying unconscious on a street and was brought to the hospital in an ambulance. He had no past history of any underlying disease but he had suffered head trauma approximately 1 month prior to this episode. On admission, he had high fever and developed a convulsion fit. Because the cerebrospinal fluid (CSF) findings showed mononuclear dominant pleocytosis and Gram staining revealed spiral-shaped gram negative bacilli, meningitis caused by C. fetus was suspected. Brain CT preformed on admission did not reveal any obvious abnormality. He was immediately treated with antibiotics effective against C. fetus. His disease was complicated by bilateral subdural empyema; therefore, bilateral burr hole drainage was performed. Durling the operation, a hematoma with an outer membrane and containing yellowish pus was revealed. Infection of a chronic subdural hematoma and consequent formation of subdural empyema was deduced. Eventually, C. fetus was isolated from the CSF arterial blood and subdural empyema. The patient was discharged with no complication after the completion of the treatment.