- Email: [email protected]
Mapping of the swine superficial lymph nodes: developing a large animal model for lymphatic interventional research
S182
Scientific e-Posters
rats underwent hepatic arterial catheterization using a left transcarotid approach and were divided into N1S1 tumorbearing (n ¼ 50) and non-tumor bearing (n ¼ 41) groups. Diameter of the proper (PH), left (LH) and right hepatic (RH) arteries, presence of anatomic variants, and animal weight were recorded at the time of angiography. Technical success of the procedure was defined as the stable placement of the microcatheter tip into the PHA. Spearman’s rank correlation coefficient (r) was calculated to determine the relationship between size of hepatic artery and weight of rats. MannWhitney and Wilcoxon tests were used to evaluate the differences between groups. Results: There was a strong positive correlation between diameter of the PH and the weight of rats in non-tumor bearing group (r ¼ 0.65; Po0.0001) and a similar moderate association was observed in the tumor bearing group (r ¼ 0.40, P ¼ 0.004).The overall success rate was 81%. There was a significant difference in both the PH diameter and the weight of rats between successful and suboptimal PH catheterization (0.59 vs 0.44mm; Po0.0001, 337.9 vs 252.6 g; Po0.0001). The left hepatic artery was significantly larger in diameter compared with the right hepatic artery (Po0.0001).Variant of anatomy was observed in one case. Conclusions: Rat weight positively correlates with proper hepatic artery diameter. Technical success rate is greater with PH diameters over 0.6 mm, typically present in animals weighing over 338 g. Familiarity with these factors can increase technical success rates of TACE in this rodent model.
Abstract No. 422 Detection of cellular and molecular markers in hepatocellular carcinoma by immunohistochemistry
Scientific e-Posters
D. Das1, S. Gilyard2, G. McLennan3, D. Das1; 1Cleveland Clinic Foundation, Cleveland, OH; 2Case Western Reserve University SOM, Cleveland, OH; 3Cleveland Clinic, Chagrin Falls, OH Purpose: The purpose of the project was to detect cellular and molecular markers using human and rat HCC tissue samples for better understanding of HCC tumor, its microenvironment and develop a realistic comparison with other solid tumors. This will greatly enhance development of better therapies in HCC. Materials: Immunohistochemistry was done for Folate receptor (FR), HIF1a, VEGF, Glial fibrillary acidic protein (GFAP), alpha smooth muscle actin (a-SMA), program death ligand 1 (PDL1) and CD11. Bright field images acquired in an Odyssey microscope and quantified using Image Pro Plus. Pixel intensity of staining and number of positive cells per high power field were counted and compared to the staining and number of positive cells in normal tissue. Results: Immunohistochemistry show increased expression of FR, HIF1a and VEGF in both rat and human HCC tissue. GFAP is a specific marker of hepatic stellate cells (HSC) in the liver. HSC are prime cell types involved in hepatic fibrosis and cirrhosis. Activated HSC synthesize a-SMA and collagen leading to remodeling of extracellular matrix and scar tissue formation. Presence of GFAP and a-SMA in HCC tissue samples, particularly tissue surrounding the tumor, confirms the presence
’
JVIR
of activated HSC in HCC–tumor microenvironment (TME). Human HCC tissue samples did not demonstrate increased expression of PDL1 but showed increased expression of CD11 indicating presence of infiltrating immune cells in HCC-TME. Conclusions: Data demonstrate that although HCC show some characteristics of other solid tumors, the HCC-TME is different in its cellular composition. HCC-TME is comprised primarily of activated HSC and infiltrating immune cells. These are important factors when targeted or immune therapy is considered for HCC treatment. Marker Normal Rats HCC Rats Normal Humans HCC Humans FR
23.2
41.4
18.38
50.00
HIF1a
17
34.5
12.25
31.5
VEGF GFAP
15.2 10.75
22.9 27.8
20.75 12.21
32.1 72.32
aSMA
2.8
29.6 12.9
24.4
CD11
Abstract No. 423 Mapping of the swine superficial lymph nodes: developing a large animal model for lymphatic interventional research D. Kraitchman1, I. Kamel2, C. Weiss3, C. Georgiades4; 1 Johns Hopkins University, Baltimore, MD; 2Johns Hopkins Hospital, Baltimore, MD; 3N/A, Baltimore, MD; 4 Johns Hopkins University, Baltimore, MD Purpose: The introduction of new lymphatic interventions has been slow, partly due to the lack of a large animal research model. Our objective was to define the percutaneously accessible, anatomically reproducible swine lymph nodes and correlate findings between MRI, ultrasound and lipiodol lymphangiography. Materials: Five adult female swine (50-60Kg) we used. Under general anesthesia, T1- and T2-weighted, coronal and axial images of the entire swine were obtained from base of skull to groins. Then, the animal’s extra-thoracic, extra-peritoneal soft tissues from the neck to the groins were scanned with ultrasound. Lymph nodes 1 cm or larger were marked before the animal was transferred to the angiography machine. Under ultrasound guidance, these nodes were accessed and lymphangiograms were obtained. The imaging findings between the three modalities were correlated and the lymph node drainage mapped. Results: Four lymph nodes/lymph node groups were identified that were reproducible in all 5 animals, larger than 1 cm and percutaneously accessible. The submandibular node, the superficial cervical lymph node group, the subiliac node and the superficial inguinal lymph node group. Drainage of these nodes mirrored human anatomy. The abdominopelvic lymphatics formed a retroperitoneal cisterna chyli and drained cephalad via a thoracic duct. Conclusions: The swine exhibits conserved lymphatic anatomy with at least four percutaneously accessible lymph nodes/node groups. Based on these results the swine may be a suitable large animal model for research into lymphatic interventions.
Scientific e-Posters
rats underwent hepatic arterial catheterization using a left transcarotid approach and were divided into N1S1 tumorbearing (n ¼ 50) and non-tumor bearing (n ¼ 41) groups. Diameter of the proper (PH), left (LH) and right hepatic (RH) arteries, presence of anatomic variants, and animal weight were recorded at the time of angiography. Technical success of the procedure was defined as the stable placement of the microcatheter tip into the PHA. Spearman’s rank correlation coefficient (r) was calculated to determine the relationship between size of hepatic artery and weight of rats. MannWhitney and Wilcoxon tests were used to evaluate the differences between groups. Results: There was a strong positive correlation between diameter of the PH and the weight of rats in non-tumor bearing group (r ¼ 0.65; Po0.0001) and a similar moderate association was observed in the tumor bearing group (r ¼ 0.40, P ¼ 0.004).The overall success rate was 81%. There was a significant difference in both the PH diameter and the weight of rats between successful and suboptimal PH catheterization (0.59 vs 0.44mm; Po0.0001, 337.9 vs 252.6 g; Po0.0001). The left hepatic artery was significantly larger in diameter compared with the right hepatic artery (Po0.0001).Variant of anatomy was observed in one case. Conclusions: Rat weight positively correlates with proper hepatic artery diameter. Technical success rate is greater with PH diameters over 0.6 mm, typically present in animals weighing over 338 g. Familiarity with these factors can increase technical success rates of TACE in this rodent model.
Abstract No. 422 Detection of cellular and molecular markers in hepatocellular carcinoma by immunohistochemistry
Scientific e-Posters
D. Das1, S. Gilyard2, G. McLennan3, D. Das1; 1Cleveland Clinic Foundation, Cleveland, OH; 2Case Western Reserve University SOM, Cleveland, OH; 3Cleveland Clinic, Chagrin Falls, OH Purpose: The purpose of the project was to detect cellular and molecular markers using human and rat HCC tissue samples for better understanding of HCC tumor, its microenvironment and develop a realistic comparison with other solid tumors. This will greatly enhance development of better therapies in HCC. Materials: Immunohistochemistry was done for Folate receptor (FR), HIF1a, VEGF, Glial fibrillary acidic protein (GFAP), alpha smooth muscle actin (a-SMA), program death ligand 1 (PDL1) and CD11. Bright field images acquired in an Odyssey microscope and quantified using Image Pro Plus. Pixel intensity of staining and number of positive cells per high power field were counted and compared to the staining and number of positive cells in normal tissue. Results: Immunohistochemistry show increased expression of FR, HIF1a and VEGF in both rat and human HCC tissue. GFAP is a specific marker of hepatic stellate cells (HSC) in the liver. HSC are prime cell types involved in hepatic fibrosis and cirrhosis. Activated HSC synthesize a-SMA and collagen leading to remodeling of extracellular matrix and scar tissue formation. Presence of GFAP and a-SMA in HCC tissue samples, particularly tissue surrounding the tumor, confirms the presence
’
JVIR
of activated HSC in HCC–tumor microenvironment (TME). Human HCC tissue samples did not demonstrate increased expression of PDL1 but showed increased expression of CD11 indicating presence of infiltrating immune cells in HCC-TME. Conclusions: Data demonstrate that although HCC show some characteristics of other solid tumors, the HCC-TME is different in its cellular composition. HCC-TME is comprised primarily of activated HSC and infiltrating immune cells. These are important factors when targeted or immune therapy is considered for HCC treatment. Marker Normal Rats HCC Rats Normal Humans HCC Humans FR
23.2
41.4
18.38
50.00
HIF1a
17
34.5
12.25
31.5
VEGF GFAP
15.2 10.75
22.9 27.8
20.75 12.21
32.1 72.32
aSMA
2.8
29.6 12.9
24.4
CD11
Abstract No. 423 Mapping of the swine superficial lymph nodes: developing a large animal model for lymphatic interventional research D. Kraitchman1, I. Kamel2, C. Weiss3, C. Georgiades4; 1 Johns Hopkins University, Baltimore, MD; 2Johns Hopkins Hospital, Baltimore, MD; 3N/A, Baltimore, MD; 4 Johns Hopkins University, Baltimore, MD Purpose: The introduction of new lymphatic interventions has been slow, partly due to the lack of a large animal research model. Our objective was to define the percutaneously accessible, anatomically reproducible swine lymph nodes and correlate findings between MRI, ultrasound and lipiodol lymphangiography. Materials: Five adult female swine (50-60Kg) we used. Under general anesthesia, T1- and T2-weighted, coronal and axial images of the entire swine were obtained from base of skull to groins. Then, the animal’s extra-thoracic, extra-peritoneal soft tissues from the neck to the groins were scanned with ultrasound. Lymph nodes 1 cm or larger were marked before the animal was transferred to the angiography machine. Under ultrasound guidance, these nodes were accessed and lymphangiograms were obtained. The imaging findings between the three modalities were correlated and the lymph node drainage mapped. Results: Four lymph nodes/lymph node groups were identified that were reproducible in all 5 animals, larger than 1 cm and percutaneously accessible. The submandibular node, the superficial cervical lymph node group, the subiliac node and the superficial inguinal lymph node group. Drainage of these nodes mirrored human anatomy. The abdominopelvic lymphatics formed a retroperitoneal cisterna chyli and drained cephalad via a thoracic duct. Conclusions: The swine exhibits conserved lymphatic anatomy with at least four percutaneously accessible lymph nodes/node groups. Based on these results the swine may be a suitable large animal model for research into lymphatic interventions.