Combined hepato-cholangiocarcinoma arising in a gallbladder intracystic papillary neoplasm. A new view on so-called "hepatoid adenocarcinoma of the gallbladder".

A 62-year-old female presented with abdominal pain, weight loss of 20 kg in the prior 6 months, and a palpable mass in the right upper quadrant during physical exam. Standard liver tests, including screening for hepatitis B and C and alpha-fetoprotein were negative or within normal limits. Computerized tomography depicted a transmural gallbladder tumor infiltrating into the adjacent liver with an irregular ill-defined mass occupying segments IV-V-VI, measuring 13.0 x 9.2 x 8.5 cm, with a solid-cystic component and heterogeneous captation of endovenous contrast media. Complete surgical resection of the neoplasm was achieved through an extended cholecystectomy and excision of hepatic segments IV, V and VI, with an uneventful follow-up 29 months until now. Morphological and immunohistochemical assessment favored a diagnosis of combined hepatocellular-cholangiocarcinoma arising in a gallbladder intracystic papillary neoplasm with invasive carcinoma. This case raises the hypothesis that the so-called "hepatoid adenocarcinoma of the gallbladder" may presently be better understood as a neoplasm derived from hepatobiliary stem/progenitor cells. Such cells have been recognized in the canals of Hering, in peribiliary glands within the liver and in the extrahepatic biliary tree, and in gallbladder mucosa.

Treatment rates in patients with chronic hepatitis C after liver biopsy.

Hepatitis C virus (HCV) infection is a major health problem in the United States. Only about 30% of patients infected with HCV are being treated despite the development of increasingly effective therapies. The aims of this study were to determine the rate of treatment for patients with HCV after undergoing liver biopsy, to assess any change in their treatment rates over recent years and to delineate the reasons for nontreatment. We retrospectively reviewed the charts of all HCV patients who had liver biopsies at Beth Israel Medical Center, New York between 1998 and 2002. The data gathered included patient demographics, stage of liver fibrosis, insurance information, treatment history and reasons for nontreatment. There were 433 liver biopsies done for chronic hepatitis C between 1998 and 2002. Of those, 267 (61%) were men. The mean age was 47 years (range, 18-72). Only 159 (37%) patients were treated after liver biopsy. Overall there were no significant differences in the treatment rates from 1999 to 2002. The common reasons for nontreatment included minimal/mild disease (stage 0-1 fibrosis, 38%), lost to follow-up or noncompliance (31%) and patient refusal (22%). Older patients more frequently had co-morbid conditions (P = 0.009). Younger age and female gender correlated with minimal disease on biopsy (P = 0.004 and 0.01, respectively). Men were lost to follow-up more frequently than women (37%vs 22%, P = 0.01). Multivariate analysis showed that age and gender were both independent predictors of minimal disease. Patients having Medicaid with or without Medicare were significantly more likely to be treated than patients with private or commercial insurance or patients with Medicare alone. A minority of HCV infected patients were treated even after having undergone liver biopsy. The proportion of HCV patients being treated after liver biopsy has been relatively stable despite advances in therapeutic success. Liver histology frequently identified patients with mild disease in whom antiviral therapy was deemed not urgent.

Postmodern biology: (adult) (stem) cells are plastic, stochastic, complex, and uncertain.

This chapter will discuss recent findings regarding cell plasticity and stem cell behavior, focusing on ways in which experimental design, observer interference, and inherent stochasticity and complexity are serving to create a new, postmodern biology. The chapter will summarize: (a) the four recognized pathways whereby cell plasticity occurs physiologically; (b) recent findings regarding unexpected epigenetic reversibility of gene restrictions that provide the mechanistic core of plasticity; (c) current evidence for the stochastic nature of gene expression and, therefore, of cell fate decisions. It will be noted that stochastic, however, does not imply completely random; rather, constrained randomness, intermediate between rigid determinism and complete disorder is what is usually seen experimentally. Possible sources of such constrained disorder, from a biomolecular point of view, will be discussed. The chapter will conclude with discussions of how these findings contribute to a Complexity Theory formulation of the body as self-organizing emergence of interacting biomolecules and the implications of such concepts for design and interpretation of experimental results (i.e., a cellular version of Heisenbergian uncertainty).

Prospects for cell-based therapies for liver disease.

Liver parenchymal maintenance and regeneration after injury are physiologically supported by 3 cell compartments: mature liver cells, intra-organ stem cells such as cells of the proximal biliary tree and periductal cells, and extra-organ stem cells from the circulation and the bone marrow. In the latter case, hepatocyte derivation from circulating cells (plasticity) can arise via direct transdifferentiation (site specific, receptor/ligand dependent) or by fusion of circulating cells with pre-existing hepatocytes. Other non-physiologic stem cells, such as mesenchymal stem cells from the bone marrow and embryonic stem cells, may be potentially used in treatment of inherited and acquired liver diseases. This review updates our current understanding of these various cell populations and of possible approaches to their future therapeutic uses in cell transplantation, bioartificial liver devices, cytokine/chemokines manipulation of physiological repair pathways, and gene therapy.

Hepatic 'stem cell' malignancies in adults: four cases.

AIMS: Combined hepatocellular/cholangiocarcinomas have been explained by some investigators as bidirectional differentiation of neoplastic progenitor cell populations. The presence of hepatic progenitor cells has now been confirmed in humans, though whether they can give rise to malignant tumours has not been confirmed. We report four cases of small tumours identified in livers with features of chronic hepatitis which may suggest a role for malignant transformation of hepatic stem cells in hepatic malignancies. METHODS: Tumour samples were studied from four patients by histochemistry and immunohistochemistry. RESULTS: Two patients had chronic hepatitis B, one had chronic hepatitis C and chronic alcoholic liver injury, and one had non-B non-C chronic hepatitis. Stages of disease ranged from portal fibrosis to cirrhosis. All tumours contained undifferentiated cells with morphological and immunohistochemical features that would be expected of hepatic progenitor cells. These cells merged with both hepatocellular carcinoma and cholangiocarcinoma components as well as with mature appearing hepatocytes within the tumours. CONCLUSION: We suggest that these tumours are of hepatic progenitor cell origin, supporting the concepts that human hepatocarcinogenesis can be based on transformation of progenitor cells and that such a process may underlie development of some mixed hepatocellular/cholangiocarcinomas and dysplastic nodules.

Toward a new paradigm of cell plasticity.

The standard paradigm of embryologic development and adult tissue reconstitution posits unidirectional, hierarchical lineages. The presumed mechanisms underlying these differentiative pathways are gene restrictions, such as methylation and heterochromatin formation, which are commonly described as irreversible. However, recent discoveries regarding multi-organ stem cells demonstrate that 'true plasticity' exists, with cells of one organ turning into cells of other organs, including differentiative transformations that cross barriers between tissues derived from different primitive germ layers. These findings, along with earlier experiments into heterokaryon formation and longstanding recognition of reactive and neoplastic lesions in humans and animals, suggest that lineage pathways are not, in fact, unidirectional. Moreover, physiologic mechanisms of reversal of gene restrictions have been recognized. Therefore, in response to these observations, we suggest a new paradigm of cell plasticity, elucidating three guiding principles of 'genomic completeness', 'uncertainty of cell characterization', and 'stochastic nature of cell origins and fates'. These principles imply a change in the way data can be interpreted and could alter subsequent hypothesis formation. This new paradigm will hopefully lead us forward to a more flexible and creative exploration of the potential of adult vertebrate cells.

Multi-organ, multi-lineage engraftment by a single bone marrow-derived stem cell.

Purification of rare hematopoietic stem cell(s) (HSC) to homogeneity is required to study their self-renewal, differentiation, phenotype, and homing. Long-term repopulation (LTR) of irradiated hosts and serial transplantation to secondary hosts represent the gold standard for demonstrating self-renewal and differentiation, the defining properties of HSC. We show that rare cells that home to bone marrow can LTR primary and secondary recipients. During the homing, CD34 and SCA-1 expression increases uniquely on cells that home to marrow. These adult bone marrow cells have tremendous differentiative capacity as they can also differentiate into epithelial cells of the liver, lung, GI tract, and skin. This finding may contribute to clinical treatment of genetic disease or tissue repair.

Hepatocellular carcinoma and dysplastic nodules in patients with cirrhosis: prospective diagnosis with MR imaging and explantation correlation.

PURPOSE: To determine the sensitivity and specificity of magnetic resonance (MR) imaging for detection of hepatocellular carcinoma (HCC) and dysplastic nodules (DNs) by using explantation correlation in patients with cirrhosis and no known HCC. MATERIALS AND METHODS: Seventy-one patients without a known history of HCC who underwent MR imaging and subsequent transplantation within 90 days were examined. Breath-hold turbo short inversion time inversion-recovery and/or T2-weighted turbo spin-echo MR images were obtained. Dynamic two- or three-dimensional gadolinium-enhanced gradient-echo MR images were obtained in the hepatic arterial, portal venous, and equilibrium phases. Prospective MR image interpretations were compared directly with explanted liver pathologic results. RESULTS: Eleven (15%) of 71 patients had hepatic malignancies; MR imaging enabled diagnosis of tumor in six (54%) of 11 patients. On a lesion-by-lesion basis, MR imaging depicted 11 of 20 hepatic neoplasms, for an overall sensitivity of 55%. MR imaging depicted four (80%) of five lesions larger than 2 cm, six (50%) of 12 lesions 1-2 cm, and one (33%) of three lesions smaller than 1 cm. MR imaging depicted only nine (15%) of 59 DNS: The specificities of MR imaging for detection of HCC and DNs on a per patient basis were 60 (86%) of 70 patients and 53 (85%) of 62 patients, respectively. CONCLUSION: MR imaging is insensitive for the diagnosis of small (<2-cm) HCCs and DNS:

Siderotic nodules in the cirrhotic liver at MR imaging with explant correlation: no increased frequency of dysplastic nodules and hepatocellular carcinoma.

PURPOSE: To determine the sensitivity of magnetic resonance (MR) imaging for detection of siderotic nodules in patients with cirrhosis and whether the frequency of hepatocellular carcinoma (HCC) and dysplastic nodules is greater if siderotic nodules are present. MATERIALS AND METHODS: MR imaging (1.5 T) was performed within 0-117 days (mean, 30 days) before liver transplantation in 77 patients. Two readers retrospectively evaluated gradient-echo (GRE) (echo time [TE], > or = 9 and 4-5 msec) and turbo short inversion time inversion-recovery or T2-weighted images for low-signal-intensity nodules. Whole-explant pathologic correlation was available in every case. RESULTS: At explantation, 28 (36%) of 77 patients had HCC, 25 (32%) had dysplastic nodules, and nine (12%) had both; 35 (45%) patients had siderotic nodules. The sensitivity of GRE imaging with 9-msec or longer TE for the detection of siderotic nodules was 80% (28 of 35) but decreased to 31% (11 of 35) with 4-5-msec TE. Frequency of HCC was not significantly higher (P =.27) in patients with (43% [15 of 35]) than in patients without (31% [13 of 42]) siderotic nodules. Frequency of dysplastic nodules also was not significantly higher (P =.42) in patients with (37% [13 of 35]) than in patients without (29% [12 of 42]) siderotic nodules. CONCLUSION: Sensitivity of MR imaging for the detection of siderotic nodules was improved with use of GRE pulse sequences with longer TEs of 9 msec or greater (80%) versus 4-5 msec (31%); however, there was no significant increased frequency of HCC or dysplastic nodules in patients with pathologically proved siderotic nodules.

Siderotic nodules at MR imaging: regenerative or dysplastic?

OBJECTIVE: To determine if iron containing "siderotic" nodules detected at magnetic resonance (MR) imaging are regenerative (RN) or dysplastic (DN) and to attempt to identify features that may distinguish them. MATERIAL AND METHODS: MR imaging (1.5 T) was performed on 77 cirrhotic patients who underwent orthotopic liver transplantation within 0-117 days (mean 30 days) of MR imaging. Two readers retrospectively evaluated breath-hold gradient-echo pulse sequences (echo time > or =9.0 ms, flip angle < or =45 degrees) for the presence of hypointense nodules, which were classified as micronodular (< or =3 mm), macronodular (>3 mm), or mixed. Nodule distribution was classified as focal (<5), scattered (5-20), or diffuse (>20) per slice. Thin section pathologic correlation was available in all cases, and Prussian blue iron stains were performed. RESULTS: Of 35 patients with pathologically proven siderotic nodules, 10 (29%) had at least 2 siderotic DN. MR detected siderotic nodules in 10 of 10 (100%) patients with siderotic DN and RN, and in 18 of 25 patients (72%) with siderotic RN only. CONCLUSION: Siderotic RN cannot be reliably distinguished from siderotic DN with MR imaging, and therefore the widely used term "siderotic regenerative nodule" should be avoided and replaced by "siderotic nodule."

Dysplastic nodules and hepatocellular carcinoma: sensitivity of digital subtraction hepatic arteriography with whole liver explant correlation.

PURPOSE: The purpose of this work was to determine the sensitivity of hepatic digital subtraction arteriography (DSA) for the detection of hepatocellular carcinoma (HCC) and dysplastic nodules (DNs) when compared with pathological findings from whole liver explants. METHOD: Twenty-one patients 30-72 years old (mean 54 years) with cirrhosis and known or clinically suspected HCC (20 prior to chemoembolization) underwent hepatic DSA with subsequent transplantation within 80 days (mean 32 days). The prospective DSA report was compared with pathologic findings from explanted livers. RESULTS: Overall, DSA detected 31 of 95 HCC lesions for a sensitivity of 33%. Of these 31 lesions, 28 were hypervascular and 3 were hypovascular. DSA detected all six HCCs measuring >5 cm, all six HCCs measuring 3-5 cm, and all five HCCs 2-3 cm, resulting in a sensitivity of 100% (17/17) for HCC >2 cm. DSA detected 7 of 18 HCCs measuring 1-2 cm (sensitivity 39%) and 7 of 60 HCCs < or =1 cm (sensitivity 12%). Overall sensitivity for DSA in detection of HCC < or =2 cm was 18% (14/78 lesions). None of 17 DNs (0.2-1.5 cm in size) was identified on DSA. CONCLUSION: DSA is insensitive to small HCC (< or =2 cm), carcinomatosis arising within nodules, and DN.

Liver from bone marrow in humans.

It has been shown in animal models that hepatocytes and cholangiocytes can derive from bone marrow cells. We have investigated whether such a process occurs in humans. Archival autopsy and biopsy liver specimens were obtained from 2 female recipients of therapeutic bone marrow transplantations with male donors and from 4 male recipients of orthotopic liver transplantations from female donors. Immunohistochemical staining with monoclonal antibody CAM5.2, specific for cytokeratins 8, 18, and 19, gave typical strong staining of hepatocytes, cholangiocytes, and ductular reactions in all tissues, to the exclusion of all nonepithelial cells. Slides were systematically photographed and then restained by fluorescence in situ hybridization (FISH) for X and Y chromosomes. Using morphologic criteria, field-by-field comparison of the fluorescent images with the prior photomicrographs, and persistence of the diaminiobenzidene (DAB) stain through the FISH protease digestion, Y-positive hepatocytes and cholangiocytes could be identified in male control liver tissue and in all study specimens. Cell counts were adjusted based on the number of Y-positive cells in the male control liver to correct for partial sampling of nuclei in the 3-micron thin tissue sections. Adjusted Y-positive hepatocyte and cholangiocyte engraftment ranged from 4% to 43% and from 4% to 38%, respectively, in study specimens, with the peak values being found in a case of fibrosing cholestatic recurrent hepatitis C in one of the liver transplant recipients. We therefore show that in humans, hepatocytes and cholangiocytes can be derived from extrahepatic circulating stem cells, probably of bone marrow origin, and such "transdifferentiation can replenish large numbers of hepatic parenchymal cells.

Focal lesions in the cirrhotic liver: high resolution ex vivo MRI with pathologic correlation.

Cirrhosis is a progressive, diffuse process of liver fibrosis that is characterized by architectural distortion and the development of a spectrum of nodules ranging from benign regenerative nodules to premalignant dysplastic nodules to overtly malignant hepatocellular carcinoma. The purpose of this essay is to demonstrate the ex vivo MR and pathology findings of these nodules as well as other masses that can be seen in the cirrhotic liver. The optimal conditions under which ex vivo imaging can be performed allow greater spatial resolution than that achieved with in vivo imaging, without artifacts that may degrade image quality. Clearly, contrast-enhanced MRI is essential for both the diagnosis and the characterization of focal lesions in the cirrhotic liver. However, the use of ex vivo imaging precludes the evaluation of these important in vivo pulse sequences.

Derivation of hepatocytes from bone marrow cells in mice after radiation-induced myeloablation.

Following a report of skeletal muscle regeneration from bone marrow cells, we investigated whether hepatocytes could also derive in vivo from bone marrow cells. A cohort of lethally irradiated B6D2F1 female mice received whole bone marrow transplants from age-matched male donors and were sacrificed at days 1, 3, 5, and 7 and months 2, 4, and 6 posttransplantation (n = 3 for each time point). Additionally, 2 archival female mice of the same strain who had previously been recipients of 200 male fluorescence-activated cell sorter (FACS)-sorted CD34(+)lin(-) cells were sacrificed 8 months posttransplantation under the same protocol. Fluorescence in situ hybridization (FISH) for the Y-chromosome was performed on liver tissue. Y-positive hepatocytes, up to 2.2% of total hepatocytes, were identified in 1 animal at 7 days posttransplantation and in all animals sacrificed 2 months or longer posttransplantation. Simultaneous FISH for the Y-chromosome and albumin messenger RNA (mRNA) confirmed male-derived cells were mature hepatocytes. These animals had received lethal doses of irradiation at the time of bone marrow transplantation, but this induced no overt, histologically demonstrable, acute hepatic injury, including inflammation, necrosis, oval cell proliferation, or scarring. We conclude that hepatocytes can derive from bone marrow cells after irradiation in the absence of severe acute injury. Also, the small subpopulation of CD34(+)lin(-) bone marrow cells is capable of such hepatic engraftment.

The canals of Hering and hepatic stem cells in humans.

Small, extraportal, hepatic parenchymal cells, positive for biliary-type cytokeratins, may represent hepatic stem cells, canals of Hering (CoH), and/or ductal plate remnants. We evaluated these cells 3 dimensionally in normal human liver and massive necrosis. Tissues from normal human livers and from 1 liver with acetaminophen-induced massive necrosis were serially sectioned, immunostained for cytokeratin 19 (CK19), and sequentially photographed. Images were examined to determine 3-dimensional relationships among CK19-positive cells. Immunostains for other hepatocyte and progenitor cell markers were examined. In normal livers, intraparenchymal CK19-positive cells lined up as linear arrays in sequential levels. One hundred of 106 (94.3%) defined, complete arrays within levels examined, most having 1 terminus at a bile duct, the other in the lobule, beyond the limiting plate. In massive necrosis, there were 767 individual CK19-positive cells or clusters around a single portal tract, 747 (97.4%) of which were spatially related forming arborizing networks connected to the interlobular bile duct by single tributaries. C-kit was positive in normal CoH. CK19 co-expressed with HepPar1, c-kit, and alpha-fetoprotein (AFP) in parenchymal cells in massive necrosis. Small, extraportal, biliary-type parenchymal cells represent cross-sections of the CoH that radiate from the portal tract, usually extending past the limiting plate into the proximate third of the hepatic lobule. The 3-dimensional structure of ductular reactions in massive necrosis suggests that these reactions are proliferations of the cells lining the CoH. Therefore, the CoH consist of, or harbor, facultative hepatic stem cells in humans.

Differential diagnosis of hepatocellular nodular lesions.

The great advances in radiologic imaging of the last two decades have focused attention on hepatic nodular lesions. Various entities with a nodular appearance are predominantly composed of hepatocytes or tumor cells of hepatocytic origin, including benign and malignant neoplasms as well as tumorlike lesions. Differential diagnosis of these nodules can often be difficult, especially in the limited material of a needle biopsy specimen. The histological features that can be of help in this regard are the focus of this review. In noncirrhotic livers, differential diagnoses include liver cell adenoma, focal nodular hyperplasia, large regenerative nodule, nodular regenerative hyperplasia, partial nodular transformation, compensatory hyperplasia, focal fatty change, and well-differentiated hepatocellular carcinoma. Poorly differentiated hepatocellular carcinoma must be distinguished from other malignant tumors, especially metastatic, poorly differentiated adenocarcinoma. In cirrhotic livers, the differential diagnoses include large regenerative nodule, focal fatty change, low-grade dysplastic nodule, high-grade dysplastic nodule, and hepatocellular carcinoma.

Dysplastic nodules in cirrhotic liver: arterial phase enhancement at CT and MR imaging--a case report.

In a 65-year-old woman with cirrhosis, biphasic helical computed tomography (CT) and multiphase gadolinium-enhanced magnetic resonance (MR) imaging depicted 11 hepatic nodules that enhanced homogeneously during the hepatic arterial phase. At pathologic examination, all lesions were dysplastic nodules with unpaired hepatic arteries within. Hepatic arterial phase enhancement of cirrhotic nodules at CT and MR imaging is not diagnostic of hepatocellular carcinoma but may occur in dysplastic nodules. Results of a single biopsy yielding hepatocellular carcinoma may not be applicable to other nodules that enhance similarly.