- Email: [email protected]
Effects of BMP-7 on liver regeneration after partial hepatectomy in mice
the second study (randotinsed, cross-over), 23 PLC (Child Pugh Score A or B) received 2 and 5 mg CLF Plasma samples were colketed at time points between 0 to 360 minutes after dosing Urine was collected pre- and post-dosing in both studies. Samples were analysed ,,wth a validated HP1C assay for C[.F. ENDPOINT8: The pharnaacokinetic (PK) parameters, including AUC, t~,~and clearance (CL) were assessed atier log-transformation. To develop a robust dynamic liver function test, several ratios of CLF plasma CL (e.g. T ~ ) were calculated In addition, a safety analysis was performed. RESULTS: In both studies AUC was proportional to the applied single iv. CLF dose, but the CL was independent of the dose administered, with a sigmficant diflerence (p<0,O01) between the mean CL of 14,8 L/h for the HVs and 8,1 L/h tot the PLC group. In addition, CL values were altered in relation to the increased Child Pugh scores and/or other markers assessing the extent of liver disease Tbe CLF T~O .~ plasma CI. ratios were in the 16 HVs with a mean of 0,32 (range: 0,21 to 0,46) significant difli:rent (p<0,O01) to the 23 PLC with a mean of 0,62 (range: 0,20 to 0,85). The reproducibility of multiple testing within individnals was high with au intra-class correlation coefficient of 0,82 tor the HVs and 0,97 for the PLC group. Furthermore, the T3c~'~ minnie CLF ratios correlated to CL and T~a, but also the severity of liver disease. The PK parameters derived from the urine showed only, minimal amorous ol CLF in the unne (up to 0,04 L&) and confirmed mainly non-renal eliminatkm mechanisms. The administration of CLF was ~t;e and well tolerated at all doses. CONCLUSION: Overall, the CLF CL showed a sigmficam and clear difference between HVs and PLC with an association to the severity of the liver disease. The CLF CL test (for example: T:~0/T~5CL ratio) is sate, reproducible and simple to pertdem in the assessment of the functional capacity of the liver. Additimta/studies are required to assess the clinical value of CLF in other liver disease conditions
(75%). Five patients with liver metastases could only be identified by CT/NMR (n=5f32; 15%). Four patients with liver metastases could only be identified by ultrasound imaging (n=4/32;12%). 2) Lesion to lesion analysis: In 18 % of the patients with liver metastases (n = 6/32) the ultrasound imaging techniques identified more metastastic lesions per individual patient than CT or NMR did In 31% of the patients with liver metastases (n = 10/32) CT or NMR identified more metastastic lesions per indi~sdual patient than the ultrasound imaging did. In 34 % of the patients with liver metastases (n = 11/32) echoenhanced ultrasound identified more metastastic lesions per individual patient than conventional b-scan sonography did. Conclusions: Echoenhanced phase inversion imaging is superior to conventional B-scan sonography in the detection of the amount of metastatic lesions per individual patient. Ultrasound imaging, even with the additional use of a echoer~hanced phase inversion imaging seems not to be able to substitute CT or NMK However, in case of a positive finding of liver metastases by" ultrasound and echoenhanced phase inversion imaging additional imaging techniques like CT and NMR could be spared.
M1439 Liver Cirrhosis Etiology by 31P MR Spectroscopy Pavel Taimr, Monika Dezortova, Milan Hajek, Julius Spicak Aims: Phosphorus magnetic resonance spectroscopy (~P MRS) enables the observation of liver metabolism in vivo through the signals of phosphomonoesters (PME), phosphodiesters (PDE), inorganic phosphate (f5) and ATP. This study deals with absolute quantificatinn of' liver metabolites obtained in 3p MR spectra of patients with fiver cirrhosis and the relationship bmween metabolite concentrations and the clinical status. Methods: 49 patients (51.4 • 10.9 yrs) with liver cirrhosis of diflerent etiology and severity described by Chlld-Pugh score (CPS) were examined afier overnight t~ting. Absolute concentrationa of PME, PDE, Pi and ATP were calculated. MR examination was pertbrmed on a whole-body MR imager Siemens Vision 1.5T using dual 3~P/~Hsurlace coil Results: Table summarizes absolute concentrations of selected metabolites together with Child-P@ score. According to ;~P MR spectra we ear* difti:rentiate vanous etiologies of cirrhosis. Alcohd liver disease (n = 19) differed in all studied metabolites (p<0.05 for Pi and ATP, p
M1437
Comparing Histological Findings with Contrast-EnhancedUhrasonography of Hepatocellular Carcinoma Nobnyuki Tatsumi, Yuji Nakayama, Gakahiro Yasuda, Tadashi Takeda, Daiki Habu, Hiroki 5akagochi, Shuhei Nishiga&i, Syuichi Seki Objectives: Contrast enhancement patterns of hepatocellular careinoina (HCC) by contrastenhanced uhramnography (CE-US) were analyeed in relation to histological differentiation of HCC. Furthermore the contrast enhancement effect was compared betveeen CE-US and dynamic CT in the early phase. Methods: The subjects of this study were 36 cases of histologically diagnosed with HCC (42 nodules m total). CE-US was performed after an intravenous bolus iqection of Levovist ~ (7 ml, 300 mg/ml). Using the FEI (flash echo imag"in-g'>colortechnique, eady phase images were taken for about 15-30 seconds, beginning 10 ~conds after the intravenous dose. A portal phase image was taken in a moment, 6090 seconds after the rejection (PowerVismn 8000, Toshiba). Dynamic CT was pertk)rmed oiler an intravmmus infusion of lopamiron ~ (100 ml, 300 mg/ml). Early phase images were taken 30 seconds alter the intusion (XVigor, Tosh:ba). The contrast enhancement pattern at each phase of CE-US was analyzed in relation to the histdogical degree of difli:rentiation of each nodular lesion of HCC The presence or absence of perfusiou (m early and portal phases was compared with that on early phase of dynamic CT Results: When examined by CE-.US, the tbllowing contrast entrancement patterns were noted. In many cases of well differenUated HCC, the tmnor was not wsible on early phase, but it svas visible on portal phase. In cases of moderately to poorly differentiated HCC, the tumor was visible on early, phase, and it remained wsible on portal phase. Thus, the contrast enhancement pattern diftered depending on the degree of tumor ditlerentiation. The perfusion positive rate on early phase was sigmficandy lower lnr nodules of well differentiated HCC (13%, 2/15) than fin" moderately difl~'rentiated HCC (75%, 12/16) and poorly differentiated HCC (75%, 6/8) (p < 0.001, X~) ~,u e~'amined by dynamic CT, the perfusion positive rate on early phase did not differ significantly between any three groups; it was 80% (12/15) for well differentiated HCC, 94% (15/16) for moderately differentiated HCC and 100% (8/8) for poorly" differentiated HCC (Table i ) Conclusion: CE-Ug can reflect the degree of tumor differentiation better than dynamic CT
Supported by" grants IGA MZ CR 6630-3 and CEZ:L17/98:00023001.
Absolute concentrations of 31P metabolttes [mM] in patients with liver ~ s | s N
r162
. . . . . w215p~O,~l
US ~ phase~ 13% (Z~15) 75% (12/16) 75~ (ee)
portal~ase 100% (15/15) 93% (14/15) ,___
Pi
POE
ATP Z70~0.94" 2.78,0,95' 2,61,0,94~ 3.60~0,94
M1440 Effects of BMP-7 on Liver Regenerationafter Partial Hepatectomy in Mice Changqing Yang, Hikam Sugimoto, Sudhakar Akulapalli, iVlauricio Giraldo, Nezam Afdhal, Michael Zeisberg, Raghu Kalluri Background: Acute liver failure is characterized by" loss of liver function and recovery., is only possible through adequate liver regeneration. TGF-hetal, a known inhibitor of prdit?ration for many kinds of cells, is also the most powerful inhibitor of hepatocyte proliferation in vitro and in vivo. Bone morphogenic protein-7 (BMP-'/), also referred to as Osteogenic protein-1 fOP-l), is a member of the TGF-betal superfamily. We have prevmusly demonstrated that BMP-7 functions as an antagonist of TGF-betal in murine models of chronic renal failure, inhibits TGF-betal induced pro-fibmtic responses. Therefore, in the present study we tested the capacity of BMP-7 to accelerate liver regeneration. Methods: 70% partial hepatectomy (PH) was performed by removal of the left and middle lobes of livers in 61 CD-1 mice. Three groups were studied; 300 mg/kg of' BMP-7 intraperitoneally (IP) ever}, other day; a placebo IP iNection and a sham surgery group. Mice were sacrificed after 2, 4, 7 and 10 days after surged~and regeneration potential was assessed by regeneration rate, tissue morphology, BrdU incorporation-proliferation index, and apoptosis using TUNEL assay. In addition TGF-hetal and hepatic growth factor (HGF) were also assessed in the tissue samples by Western blot and immunohistochemistry. Liver function was analyzed by serum ALT, AST and total bilirubin measurement. Results: Systemic admimstmnon of BMP~ 7 lead to a dramatic acceleration of liver regeneration at the groups of 2 days, 4 days and 7 days after PH, according to the BrdU index and liver regeneration rate, compared with control group and sham group Hepatocytes, which can express ALK-3 (type I receptor of BMP-7), become the tnain source of TGF-betal after PH. The expression HGF was not affected by the administration of BMP-7, however, the increase of TGF-betal expression afier PH was do,an-regulated. Apoptosis of hepatocytes also decreased at 7 day~ and 10days after PH in the BMP-7 treated groups. Liver tu.ncrion was improved at 4 days and 7 days after PH in the treated group. Conclusion: BMP-7 is an eftective nomtoxic protein capable of significant acceleration of liver regeneration BMP-7 appears to promote regeneration by down regulation of TGF-betal and by reducing hepatocyte apoptosis. Further studies are needed to evaluate a potential clinical role for BMP-7 m acute liver failure.
Perfuston positive rates for different imaging techniques hiMololgy well moderately
PME
All pat~erR~ 49 3.06~1.18 1.22~0.58" 5,32,2.64 s CPS: A+B 26 3,15 L06 1 23~0,59" 6.95 24Z -~ CPS:C 23 295~1,32 1.20~0,5T 5.67 2.75~,* Conbx/~ 9 2.78~1.27 1.690.70 9,88~2.69 *)p<005, s)p<0.01,~p<0.001from controls; *}p<0.05between CPS A+B to CPS: C
dynamic CT early phase 80% (12/~5) 94% (15116)
100~ ( ~ ) ............................
M1438 Detection of Liver Metastasis by Echo-EnhancedUltrasound - Interim Analysis of Prospective Multicenter Trial D Strobel, I". Bernatik, S. Heller, A. Bunk~ K Hi~chbuehl, G Adler, E G. Hahn, W. Kratzer Aim: To compare the diagnostic accuracy of echoenhanced ultrasound tor the detection of liver metastases in comparison to computed tomography, and nJagnetic resonance imagmg in a ongoing prospective multicenter trial Methods: Patients with known extrahepatic ~r~alignancy were screened tor lwer metastases using conventional B-scan sonography and echoenhanced ultrasouud (phase inversion imaging, Souovue 2.4 ml i.v. Bolus, Altana Pharma, Konstanz, Germany). The detectinn rate of liver metastases was compared to biphasic computed tomogaphy (CT) and maguetic resonance imaging (NMR). hi addition a lesion by lesion comparison was performed. Liver metastases was confirmed after a 3-6 months follow up. Resuhs: Up to nuw 46 patients with colorectal cancer (n = 20), pancreatic cancer (n = 13), gastric cancer (n = 4), breast cancer (n = 3), esophageal cancer (n = 1), lung cancer (n = I), cholangiocarcinoma (n = I), malignant melanoma (n= 1), adrenal cancer (n= 1), caminoid tumor (n = 1) were analyzed, in 32 patients liver metastases was identified: t) Detection late of hver metastases: CT/NMR detected liver metastases in 28/32 patients (87%) compared to B-scan-sonography/ecfio-enhanced pbase inversion imaging in 24/32 patients
A-755
AASLD
Abstracts
(75%). Five patients with liver metastases could only be identified by CT/NMR (n=5f32; 15%). Four patients with liver metastases could only be identified by ultrasound imaging (n=4/32;12%). 2) Lesion to lesion analysis: In 18 % of the patients with liver metastases (n = 6/32) the ultrasound imaging techniques identified more metastastic lesions per individual patient than CT or NMR did In 31% of the patients with liver metastases (n = 10/32) CT or NMR identified more metastastic lesions per indi~sdual patient than the ultrasound imaging did. In 34 % of the patients with liver metastases (n = 11/32) echoenhanced ultrasound identified more metastastic lesions per individual patient than conventional b-scan sonography did. Conclusions: Echoenhanced phase inversion imaging is superior to conventional B-scan sonography in the detection of the amount of metastatic lesions per individual patient. Ultrasound imaging, even with the additional use of a echoer~hanced phase inversion imaging seems not to be able to substitute CT or NMK However, in case of a positive finding of liver metastases by" ultrasound and echoenhanced phase inversion imaging additional imaging techniques like CT and NMR could be spared.
M1439 Liver Cirrhosis Etiology by 31P MR Spectroscopy Pavel Taimr, Monika Dezortova, Milan Hajek, Julius Spicak Aims: Phosphorus magnetic resonance spectroscopy (~P MRS) enables the observation of liver metabolism in vivo through the signals of phosphomonoesters (PME), phosphodiesters (PDE), inorganic phosphate (f5) and ATP. This study deals with absolute quantificatinn of' liver metabolites obtained in 3p MR spectra of patients with fiver cirrhosis and the relationship bmween metabolite concentrations and the clinical status. Methods: 49 patients (51.4 • 10.9 yrs) with liver cirrhosis of diflerent etiology and severity described by Chlld-Pugh score (CPS) were examined afier overnight t~ting. Absolute concentrationa of PME, PDE, Pi and ATP were calculated. MR examination was pertbrmed on a whole-body MR imager Siemens Vision 1.5T using dual 3~P/~Hsurlace coil Results: Table summarizes absolute concentrations of selected metabolites together with Child-P@ score. According to ;~P MR spectra we ear* difti:rentiate vanous etiologies of cirrhosis. Alcohd liver disease (n = 19) differed in all studied metabolites (p<0.05 for Pi and ATP, p
M1437
Comparing Histological Findings with Contrast-EnhancedUhrasonography of Hepatocellular Carcinoma Nobnyuki Tatsumi, Yuji Nakayama, Gakahiro Yasuda, Tadashi Takeda, Daiki Habu, Hiroki 5akagochi, Shuhei Nishiga&i, Syuichi Seki Objectives: Contrast enhancement patterns of hepatocellular careinoina (HCC) by contrastenhanced uhramnography (CE-US) were analyeed in relation to histological differentiation of HCC. Furthermore the contrast enhancement effect was compared betveeen CE-US and dynamic CT in the early phase. Methods: The subjects of this study were 36 cases of histologically diagnosed with HCC (42 nodules m total). CE-US was performed after an intravenous bolus iqection of Levovist ~ (7 ml, 300 mg/ml). Using the FEI (flash echo imag"in-g'>colortechnique, eady phase images were taken for about 15-30 seconds, beginning 10 ~conds after the intravenous dose. A portal phase image was taken in a moment, 6090 seconds after the rejection (PowerVismn 8000, Toshiba). Dynamic CT was pertk)rmed oiler an intravmmus infusion of lopamiron ~ (100 ml, 300 mg/ml). Early phase images were taken 30 seconds alter the intusion (XVigor, Tosh:ba). The contrast enhancement pattern at each phase of CE-US was analyzed in relation to the histdogical degree of difli:rentiation of each nodular lesion of HCC The presence or absence of perfusiou (m early and portal phases was compared with that on early phase of dynamic CT Results: When examined by CE-.US, the tbllowing contrast entrancement patterns were noted. In many cases of well differenUated HCC, the tmnor was not wsible on early phase, but it svas visible on portal phase. In cases of moderately to poorly differentiated HCC, the tumor was visible on early, phase, and it remained wsible on portal phase. Thus, the contrast enhancement pattern diftered depending on the degree of tumor ditlerentiation. The perfusion positive rate on early phase was sigmficandy lower lnr nodules of well differentiated HCC (13%, 2/15) than fin" moderately difl~'rentiated HCC (75%, 12/16) and poorly differentiated HCC (75%, 6/8) (p < 0.001, X~) ~,u e~'amined by dynamic CT, the perfusion positive rate on early phase did not differ significantly between any three groups; it was 80% (12/15) for well differentiated HCC, 94% (15/16) for moderately differentiated HCC and 100% (8/8) for poorly" differentiated HCC (Table i ) Conclusion: CE-Ug can reflect the degree of tumor differentiation better than dynamic CT
Supported by" grants IGA MZ CR 6630-3 and CEZ:L17/98:00023001.
Absolute concentrations of 31P metabolttes [mM] in patients with liver ~ s | s N
r162
. . . . . w215p~O,~l
US ~ phase~ 13% (Z~15) 75% (12/16) 75~ (ee)
portal~ase 100% (15/15) 93% (14/15) ,___
Pi
POE
ATP Z70~0.94" 2.78,0,95' 2,61,0,94~ 3.60~0,94
M1440 Effects of BMP-7 on Liver Regenerationafter Partial Hepatectomy in Mice Changqing Yang, Hikam Sugimoto, Sudhakar Akulapalli, iVlauricio Giraldo, Nezam Afdhal, Michael Zeisberg, Raghu Kalluri Background: Acute liver failure is characterized by" loss of liver function and recovery., is only possible through adequate liver regeneration. TGF-hetal, a known inhibitor of prdit?ration for many kinds of cells, is also the most powerful inhibitor of hepatocyte proliferation in vitro and in vivo. Bone morphogenic protein-7 (BMP-'/), also referred to as Osteogenic protein-1 fOP-l), is a member of the TGF-betal superfamily. We have prevmusly demonstrated that BMP-7 functions as an antagonist of TGF-betal in murine models of chronic renal failure, inhibits TGF-betal induced pro-fibmtic responses. Therefore, in the present study we tested the capacity of BMP-7 to accelerate liver regeneration. Methods: 70% partial hepatectomy (PH) was performed by removal of the left and middle lobes of livers in 61 CD-1 mice. Three groups were studied; 300 mg/kg of' BMP-7 intraperitoneally (IP) ever}, other day; a placebo IP iNection and a sham surgery group. Mice were sacrificed after 2, 4, 7 and 10 days after surged~and regeneration potential was assessed by regeneration rate, tissue morphology, BrdU incorporation-proliferation index, and apoptosis using TUNEL assay. In addition TGF-hetal and hepatic growth factor (HGF) were also assessed in the tissue samples by Western blot and immunohistochemistry. Liver function was analyzed by serum ALT, AST and total bilirubin measurement. Results: Systemic admimstmnon of BMP~ 7 lead to a dramatic acceleration of liver regeneration at the groups of 2 days, 4 days and 7 days after PH, according to the BrdU index and liver regeneration rate, compared with control group and sham group Hepatocytes, which can express ALK-3 (type I receptor of BMP-7), become the tnain source of TGF-betal after PH. The expression HGF was not affected by the administration of BMP-7, however, the increase of TGF-betal expression afier PH was do,an-regulated. Apoptosis of hepatocytes also decreased at 7 day~ and 10days after PH in the BMP-7 treated groups. Liver tu.ncrion was improved at 4 days and 7 days after PH in the treated group. Conclusion: BMP-7 is an eftective nomtoxic protein capable of significant acceleration of liver regeneration BMP-7 appears to promote regeneration by down regulation of TGF-betal and by reducing hepatocyte apoptosis. Further studies are needed to evaluate a potential clinical role for BMP-7 m acute liver failure.
Perfuston positive rates for different imaging techniques hiMololgy well moderately
PME
All pat~erR~ 49 3.06~1.18 1.22~0.58" 5,32,2.64 s CPS: A+B 26 3,15 L06 1 23~0,59" 6.95 24Z -~ CPS:C 23 295~1,32 1.20~0,5T 5.67 2.75~,* Conbx/~ 9 2.78~1.27 1.690.70 9,88~2.69 *)p<005, s)p<0.01,~p<0.001from controls; *}p<0.05between CPS A+B to CPS: C
dynamic CT early phase 80% (12/~5) 94% (15116)
100~ ( ~ ) ............................
M1438 Detection of Liver Metastasis by Echo-EnhancedUltrasound - Interim Analysis of Prospective Multicenter Trial D Strobel, I". Bernatik, S. Heller, A. Bunk~ K Hi~chbuehl, G Adler, E G. Hahn, W. Kratzer Aim: To compare the diagnostic accuracy of echoenhanced ultrasound tor the detection of liver metastases in comparison to computed tomography, and nJagnetic resonance imagmg in a ongoing prospective multicenter trial Methods: Patients with known extrahepatic ~r~alignancy were screened tor lwer metastases using conventional B-scan sonography and echoenhanced ultrasouud (phase inversion imaging, Souovue 2.4 ml i.v. Bolus, Altana Pharma, Konstanz, Germany). The detectinn rate of liver metastases was compared to biphasic computed tomogaphy (CT) and maguetic resonance imaging (NMR). hi addition a lesion by lesion comparison was performed. Liver metastases was confirmed after a 3-6 months follow up. Resuhs: Up to nuw 46 patients with colorectal cancer (n = 20), pancreatic cancer (n = 13), gastric cancer (n = 4), breast cancer (n = 3), esophageal cancer (n = 1), lung cancer (n = I), cholangiocarcinoma (n = I), malignant melanoma (n= 1), adrenal cancer (n= 1), caminoid tumor (n = 1) were analyzed, in 32 patients liver metastases was identified: t) Detection late of hver metastases: CT/NMR detected liver metastases in 28/32 patients (87%) compared to B-scan-sonography/ecfio-enhanced pbase inversion imaging in 24/32 patients
A-755
AASLD
Abstracts