The efficacy of an AKT inhibitor for treatment of aggressive endometrial cancer: Studied in new...
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were treated with 30.4 μM medroxyprogesterone 17-acetate (MPA) for 6 days, and cell counts, cell cycle analysis, apoptosis assays, and flow cytometry were performed to analyze the effects of MPA treatment. Expression of estrogen (ER), progesterone (PR) and androgen (AR) receptors was demonstrated by western blot and immunocytochemistry (ICC). T-test with p-values b 0.05 was used to determine statistical sig- nificance between groups. Results: The proportions of CD133 + cells in Ishikawa (80.7%) and KLE (6.5%) cell lines were determined. After MPA treatment, there were significant reductions in the percentage of total live cells when compared to the controls in Ishikawa (86.4% vs. 100%) and KLE (45.6% vs. 100%) cell lines. Additionally, statistically significant reductions in CD133+ cells with MPA were present when compared to non-treated controls in Ishikawa (62.2% vs. 81.1%) and KLE (2.7 vs. 6.4%) cell lines. A significant increase in the presence of apoptotic cells was noted in all both lines treated with MPA compared to controls (Ishikawa 32.1% vs. 0.4%; KLE 28.4% vs. 0.7%). In both the Ishikawa and KLE lines, there was no difference in the percent of cells in the S + G2M phases of cell cycle. Western blots and ICC demonstrated the presence of ER and PR in only the Ishikawa line. AR was present in Ishikawa and KLE. Conclusion: In Ishikawa and KLE endometrial cancer cell lines, the multifactoral effects of high dose progestins are to decrease the CD133+ stem cells, decrease total cell counts through upregulation of apoptotis. These effects were also noted in ER/PR negative cell line that expressed AR. In addition to traditional PR signaling pathways, off-target AR signaling may be the more relevant modulator of stem cell control. doi:10.1016/j.ygyno.2014.07.033 The efficacy of an AKT inhibitor for treatment of aggressive endometrial cancer: Studied in new models for cancer research A. Winder, Y. Yu, K. Unno, J.J. Kim. Northwestern University, Chicago, IL, USA. Objectives: Data is accumulating that type II endometrial adenocar- cinomas including serous, clear cell, and endometrioid histologies share cellular pathway aberrations responsible for a unique molec- ular landscape. PIK3CA is commonly mutated in these aggressive cancers, leading to a constitutively activated PI3K/AKT pathway and subsequent tumor growth. This pathway may represent a novel therapeutic target. The purpose of this study was to compare tumor growth and the invasive behavior of aggressive uterine adenocarci- nomas treated with an AKT inhibitor. Methods: Using primary patient derived tumor isolates with histologically-confirmed, type II uterine cancer and immunohisto- chemistry proven over-expression of activated AKT, we examined tumor growth in response to an AKT inhibitor, MK2206, in a renal capsule mouse xenograft model, 3D polystyrene scaffold, and 2D cell culture. We also used the AN3CA cell line that was derived from a uterine adenocarcinoma lymph node metastasis with a homozygous PTEN mutation. Tumor was placed under the renal capsule of NOD scid gamma (NSG) mice that were treated with MK2206 or vehicle control and tumor volume was measured. Patient isolates and AN3CA cells were grown in 3D culture, treated with MK2206, and examined with H&E staining. These same cells were treated with increasing concentrations of MK2206 in 2D cell culture; levels of activated AKT, total AKT, and activated PRAS were compared by western blot. The invasive behavior of these cells was evaluated using a matrigel invasion chamber assay in response to MK2206 compared to vehicle control. Results: Tumor growth of patient derived tissues was notably inhibited in the NSG mice treated with oral MK2206 compared to vehicle control. There was more than a 2 fold decrease in tumor volume in the group treated with MK2206 compared to the control. All the mice survived with minimal toxicity. 3D cell culture is a new platform for cancer investigation and we grew both the AN3CA cell line and patient derived tumor isolates in this model. Additionally, we found an effective dose-dependent decrease in activated AKT in both patient tumor cells and the AN3CA cells when treated with MK2206 in 2D culture. Regarding invasion, the number of the patient tumor cells and AN3CA cells invading the matrigel inserts was greatly decreased by 1 μM MK2206. Conclusion: Current treatment of aggressive uterine cancer is poor. These novel models show that targeting the PI3K/AKT pathway reduces tumor growth and minimizes invasion. doi:10.1016/j.ygyno.2014.07.034 Surgical complications following robotically-assisted gynecologic oncology procedures A. Barrie, C. Garcia, A.H. Freeman, R.D. Littell, B. Powell. Gynecologic Oncology, Kaiser Permanente San Francisco Medical Center, CA, USA. Objectives: Although surgical complications are well described among laparoscopic and open oncologic procedures, these compli- cations following robotic surgery are less understood. This study aims to evaluate surgical complications of robotically-assisted laparo- scopic surgeries. Methods: Women undergoing robotically-assisted gynecologic on- cology surgery from December 2008 to January 2014 were identified from a large community based integrated health system in Northern California. Patients with incomplete records for review were ex- cluded from analysis. A retrospective chart review using the elec- tronic medical record was performed. Primary outcome was the incidence of surgical complications within 30 days following the procedure. Minor complications were classified as any deviation from normal postoperative course with or without need for pharma- cological treatment, including blood transfusions and total parenteral nutrition. Major complications were classified as requiring surgical, endoscopic, radiologic intervention under general or local anesthesia, life threatening complications, CNS complication, single organ dys- function, multi-organ failure, and death. We stratified based on the Clavien–Dindo surgical complication classification, with grades 1–2 as minor and grades 3–5 as major. Results: A total number of 1110 cases were identified from December 2008 to January 2014. As data is collected from an integrated health care system, all patient information and follow-up are captured in an electronic record, thus few patients are lost to follow-up and complete data is available on all complications. Ten patients were excluded due to incomplete medical records. Among these cases, we identified 357 total complications: 108 (9.7%) were major and 249 (22.4%) were minor complications. Major complica- tions were more common in radical trachelectomies and radical parametrectomies compared with hysterectomy, nodal dissections and radical hysterectomy. Women undergoing robotically-assisted gynecology oncology procedure were found to have the following rates of complications in the table below. Overall rates of major complications included ureteral injury (10, 0.01%), fistula (3, 0.003%), bowel injury (3, 0.002%), vascular injury (1, 0.001%), pulmonary embolus (4, 0.003%), abscess (16, 0.014%), and death (2, 0.002%). Conclusion: As we continue to refine robotic surgery, further studies are to directly compare the risks between laparoscopy and robotically assisted surgery in this cohort. The rates of major complications (9.7%) can be compared to rates of moderate and severe complications from the GOG Lap2 study (14%), demonstrating a possible reduction in complications with robotic surgery. An electronic data collection Abstracts 392
Transcript of The efficacy of an AKT inhibitor for treatment of aggressive endometrial cancer: Studied in new...
were treated with 30.4 μM medroxyprogesterone 17-acetate (MPA)for 6 days, and cell counts, cell cycle analysis, apoptosis assays, and flowcytometry were performed to analyze the effects of MPA treatment.Expression of estrogen (ER), progesterone (PR) and androgen (AR)receptors was demonstrated bywestern blot and immunocytochemistry(ICC). T-test with p-values b0.05 was used to determine statistical sig-nificance between groups.Results: The proportions of CD133+ cells in Ishikawa (80.7%)and KLE (6.5%) cell lines were determined. After MPA treatment,there were significant reductions in the percentage of total live cellswhen compared to the controls in Ishikawa (86.4% vs. 100%) andKLE (45.6% vs. 100%) cell lines. Additionally, statistically significantreductions in CD133+ cells with MPA were present when comparedto non-treated controls in Ishikawa (62.2% vs. 81.1%) and KLE (2.7 vs.6.4%) cell lines. A significant increase in the presence of apoptoticcells was noted in all both lines treated with MPA compared tocontrols (Ishikawa 32.1% vs. 0.4%; KLE 28.4% vs. 0.7%). In both theIshikawa and KLE lines, there was no difference in the percent ofcells in the S + G2M phases of cell cycle. Western blots and ICCdemonstrated the presence of ER and PR in only the Ishikawa line.AR was present in Ishikawa and KLE.Conclusion: In Ishikawa and KLE endometrial cancer cell lines, themultifactoral effects of high dose progestins are to decrease theCD133+ stem cells, decrease total cell counts through upregulationof apoptotis. These effects were also noted in ER/PR negative cell linethat expressed AR. In addition to traditional PR signaling pathways,off-target AR signaling may be the more relevant modulator of stemcell control.
doi:10.1016/j.ygyno.2014.07.033
The efficacy of an AKT inhibitor for treatment of aggressiveendometrial cancer: Studied in new models for cancer researchA. Winder, Y. Yu, K. Unno, J.J. Kim. Northwestern University, Chicago,IL, USA.
Objectives: Data is accumulating that type II endometrial adenocar-cinomas including serous, clear cell, and endometrioid histologiesshare cellular pathway aberrations responsible for a unique molec-ular landscape. PIK3CA is commonly mutated in these aggressivecancers, leading to a constitutively activated PI3K/AKT pathway andsubsequent tumor growth. This pathway may represent a noveltherapeutic target. The purpose of this study was to compare tumorgrowth and the invasive behavior of aggressive uterine adenocarci-nomas treated with an AKT inhibitor.Methods: Using primary patient derived tumor isolates withhistologically-confirmed, type II uterine cancer and immunohisto-chemistry proven over-expression of activated AKT, we examinedtumor growth in response to an AKT inhibitor, MK2206, in a renalcapsule mouse xenograft model, 3D polystyrene scaffold, and 2D cellculture. We also used the AN3CA cell line that was derived from auterine adenocarcinoma lymph node metastasis with a homozygousPTEN mutation. Tumor was placed under the renal capsule of NODscid gamma (NSG) mice that were treated with MK2206 or vehiclecontrol and tumor volume was measured. Patient isolates and AN3CAcells were grown in 3D culture, treated with MK2206, and examinedwith H&E staining. These same cells were treated with increasingconcentrations of MK2206 in 2D cell culture; levels of activated AKT,total AKT, and activated PRAS were compared by western blot. Theinvasive behavior of these cells was evaluated using a matrigelinvasion chamber assay in response to MK2206 compared to vehiclecontrol.Results: Tumor growth of patient derived tissues was notablyinhibited in the NSG mice treated with oral MK2206 compared to
vehicle control. There was more than a 2 fold decrease in tumorvolume in the group treated with MK2206 compared to the control.All the mice survived with minimal toxicity. 3D cell culture is a newplatform for cancer investigation and we grew both the AN3CA cellline and patient derived tumor isolates in this model. Additionally,we found an effective dose-dependent decrease in activated AKT inboth patient tumor cells and the AN3CA cells when treated withMK2206 in 2D culture. Regarding invasion, the number of the patienttumor cells and AN3CA cells invading the matrigel inserts wasgreatly decreased by 1 μM MK2206.Conclusion: Current treatment of aggressive uterine cancer is poor.These novel models show that targeting the PI3K/AKT pathway reducestumor growth and minimizes invasion.
doi:10.1016/j.ygyno.2014.07.034
Surgical complications following robotically-assisted gynecologiconcology proceduresA. Barrie, C. Garcia, A.H. Freeman, R.D. Littell, B. Powell. GynecologicOncology, Kaiser Permanente San Francisco Medical Center, CA, USA.
Objectives: Although surgical complications are well describedamong laparoscopic and open oncologic procedures, these compli-cations following robotic surgery are less understood. This studyaims to evaluate surgical complications of robotically-assisted laparo-scopic surgeries.Methods: Women undergoing robotically-assisted gynecologic on-cology surgery from December 2008 to January 2014 were identifiedfrom a large community based integrated health system in NorthernCalifornia. Patients with incomplete records for review were ex-cluded from analysis. A retrospective chart review using the elec-tronic medical record was performed. Primary outcome was theincidence of surgical complications within 30 days following theprocedure. Minor complications were classified as any deviationfrom normal postoperative course with or without need for pharma-cological treatment, including blood transfusions and total parenteralnutrition. Major complications were classified as requiring surgical,endoscopic, radiologic intervention under general or local anesthesia,life threatening complications, CNS complication, single organ dys-function, multi-organ failure, and death. We stratified based on theClavien–Dindo surgical complication classification, with grades 1–2as minor and grades 3–5 as major.Results: A total number of 1110 cases were identified fromDecember 2008 to January 2014. As data is collected from anintegrated health care system, all patient information and follow-upare captured in an electronic record, thus few patients are lost tofollow-up and complete data is available on all complications. Tenpatients were excluded due to incomplete medical records. Amongthese cases, we identified 357 total complications: 108 (9.7%) weremajor and 249 (22.4%) were minor complications. Major complica-tions were more common in radical trachelectomies and radicalparametrectomies compared with hysterectomy, nodal dissectionsand radical hysterectomy. Women undergoing robotically-assistedgynecology oncology procedure were found to have the followingrates of complications in the table below. Overall rates of majorcomplications included ureteral injury (10, 0.01%), fistula (3, 0.003%),bowel injury (3, 0.002%), vascular injury (1, 0.001%), pulmonaryembolus (4, 0.003%), abscess (16, 0.014%), and death (2, 0.002%).Conclusion: As we continue to refine robotic surgery, further studiesare to directly compare the risks between laparoscopy and roboticallyassisted surgery in this cohort. The rates ofmajor complications (9.7%)can be compared to rates of moderate and severe complications fromthe GOG Lap2 study (14%), demonstrating a possible reduction incomplications with robotic surgery. An electronic data collection
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