Anna Gukovskaya

University of California, Los Angeles | UCLA

Intraperitoneal administration of high doses of basic amino acids, such as L-lysine (L-Lys), L-arginine (L-Arg) or L-ornithine (L-Orn) induces acute pancreatitis in rodents. Although the exact mechanism of their action is not fully understood, the role of mitochondria has been implicated. We aimed to investigate the effects of basic amino acids, pa...

Pancreatitis is a potentially fatal disease of the exocrine pancreas and one of the most common reasons for hospital admissions for those with GI disease [1–5]. The etiology of human pancreatitis includes gallstones, alcohol abuse, and the effect of various genetic abnormalities. There are two major forms of the disease, acute and chronic [1–11]. S...

Excessive alcohol intake is a major risk factor for pancreatitis, sensitizing exocrine pancreas to stressors by mechanisms that remain obscure. Impaired autophagy drives non-alcoholic pancreatitis, but the effects of ethanol (EtOH) and alcoholic pancreatitis on autophagy are poorly understood. Here, we find that ethanol reduces autophagosome format...

Background & Aims: Inflammation is a major determinant of the severity of acute pancreatitis (AP); however, there are no treatments currently available to reduce persistent/uncontrolled inflammation and associated morbidity of pancreatitis. We hypothesized that the interplay between key transcription factor pathways, the proinflammatory NF-κB and t...

Background and Aims: Dysfunction of acinar cell lysosomal and mitochondrial machinery initiates and drives pancreatitis. However, the underlying mechanisms remain poorly understood, and interrelationships between lysosomal and mitochondrial dysfunctions in pancreatitis have not been explored. Furthermore, approaches to normalize lysosomal/autophagy...

Background and Aims: Cholesterol is a critical component of cell membranes regulating multiple organelle functions. Our recent study revealed that acinar cell cholesterol homeostasis is severely disrupted in pancreatitis. Here we examine the mechanisms of cholesterol disordering in pancreatitis, particularly the role of the key lysosomal cholestero...

Background Autophagosome, the central organelle in autophagy process, can assemble via canonical pathway mediated by LC3-II, the lipidated form of autophagy-related protein LC3/ATG8, or noncanonical pathway mediated by the small GTPase Rab9. Canonical autophagy is essential for exocrine pancreas homeostasis, and its disordering initiates and drives...

Disordered lysosomal/autophagy pathways initiate and drive pancreatitis, but the underlying mechanisms and links to disease pathology are poorly understood. Here, we show that mannose-6-phosphate (M6P) pathway of hydrolase delivery to lysosomes critically regulates pancreatic acinar cell cholesterol metabolism. Ablation of the Gnptab gene coding fo...

Acute pancreatitis (AP), an acute inflammatory disorder of the exocrine pancreas, is one of the most common gastrointestinal diseases encountered in emergency departments with no specific treatments. Laboratory-based research has formed the cornerstone of endeavours to decipher the pathophysiology of AP, because of the limitations of such study in...

Premature intrapancreatic trypsinogen activation is widely regarded as an initiating event for acute pancreatitis. Previous studies have alternatively implicated secretory vesicles, endosomes, lysosomes, or autophagosomes/autophagolysosomes as the primary site of trypsinogen activation, from which a cell-damaging proteolytic cascade originates. To...

Pancreatitis is a common, sometimes fatal, disease of exocrine pancreas, initiated by damaged acinar cells. Recent studies implicate disordered macroautophagy/autophagy in pancreatitis pathogenesis. ATG8/LC3 protein is critical for autophagosome formation and a widely used marker of autophagic vacuoles. Transgenic GFP-LC3 mice are a valuable tool t...

Acute pancreatitis (AP) is a potentially lethal inflammatory disease that lacks specific therapy. Damaged pancreatic acinar cells are believed to be the site of AP initiation. The primary function of these cells is the synthesis, storage, and export of digestive enzymes. Beginning in the endoplasmic reticulum and ending with secretion of proteins s...

Impaired macroautophagy/autophagy has been implicated in experimental and human pancreatitis. However, the transcriptional control governing the autophagy-lysosomal process in pancreatitis is largely unknown. We investigated the role and mechanisms of TFEB (transcription factor EB), a master regulator of lysosomal biogenesis, in the pathogenesis of...

Objective: The role of autophagy in pancreatic neoplasia is controversial, with reports supporting both tumor promoting and suppressing functions. Mechanisms underlying these opposing effects are poorly understood. Here, we evaluate the role of Beclin1, a key autophagy mediator, in the development of pancreatic intraepithelial neoplasia (PanIN) in...

Background & Aims Despite accumulating evidence for important role of autophagy in pancreatitis, the underlying mechanisms are poorly understood. Conversion of the key autophagy‐related (Atg) protein Atg8/LC3 fromits cytosolic LC3‐I form to the lipidated LC3‐II form is a critical step in autophagosome formation. We used mice (over)expressing LC3 co...

Background & aims: The pathogenic mechanism of pancreatitis is poorly understood. Recent evidence implicates defective autophagy in pancreatitis responses; however, the pathways mediating impaired autophagy in pancreas remain largely unknown. Here, we investigate the role of lysosome associated membrane proteins (LAMPs) in pancreatitis. Methods:...

Acute pancreatitis is caused by toxins that induce acinar cell calcium overload, zymogen activation, cytokine release and cell death, yet is without specific drug therapy. Mitochondrial dysfunction has been implicated but the mechanism not established. We investigated the mechanism of induction and consequences of the mitochondrial permeability tra...

A few years ago, autophagy would require a detailed background introduction, but more and more, this fundamental cell biology mechanism of lysosome-driven degradation and recycling of cellular organelles, long-lived proteins, and lipids is becoming a common knowledge. Its main pathway, macroautophagy (herein referred to as autophagy), sequesters ma...

Introduction: There is strong epidemiologic evidence that obesity increases the risk of several human cancers, including pancreatic cancer. Several mechanisms, including inflammation and insulin resistance with hyperinsulinemia, are proposed, by which obesity may promote tumor development. However, the driving mechanisms in pancreatic cancer are st...

Research in autophagy continues to accelerate,(1) and as a result many new scientists are entering the field. Accordingly, it is important to establish a standard set of criteria for monitoring macroautophagy in different organisms. Recent reviews have described the range of assays that have been used for this purpose.(2,3) There are many useful an...

229 Background: Chronic pancreatitis (CP) and smoking are two major risk factors for pancreatic cancer (PaCa). Here we developed mouse models of PaCa by exposing mice to CP and/or smoking. Methods: Wild type (WT) and Pdx1-Cre;LSL-Kras transgenic mice were subjected to 7-hourly cerulein injections (50µg/kg) twice a week for 3 weeks to induce CP. Ano...

Proceedings: AACR 104th Annual Meeting 2013; Apr 6-10, 2013; Washington, DC Background and Aims: Smoking and pancreatitis are two major risk factors for pancreatic cancer. We developed novel mouse models of pancreatic cancer induced by smoking and chronic pancreatitis. We used two pancreatic cancer mouse models induced by overexpression of Kras wi...

There is epidemiologic evidence that obesity increases the risk of cancers. Several underlying mechanisms, including inflammation and insulin resistance, are proposed. However, the driving mechanisms in pancreatic cancer are poorly understood. The goal of the present study was to develop a model of diet-induced obesity and pancreatic cancer develop...

Inflammation and autophagy are cellular defense mechanisms. When these processes are deregulated (deficient or overactivated) they produce pathologic effects, such as oxidative stress, metabolic impairments, and cell death. Unresolved inflammation and disrupted regulation of autophagy are common features of pancreatitis and pancreatic cancer. Furth...

Chronic pancreatitis is an inflammatory disease that causes progressive destruction of pancreatic acinar cells and, ultimately, loss of pancreatic function. We investigated the role of IκB kinase α (IKKα) in pancreatic homeostasis. Pancreas-specific ablation of IKKα (IkkαΔpan) caused spontaneous and progressive acinar cell vacuolization and death,...

The pathogenic mechanisms underlying acute pancreatitis are not clear. Two key pathologic acinar cell responses of this disease are vacuole accumulation and trypsinogen activation. We show here that both result from defective autophagy, by comparing the autophagic responses in rodent models of acute pancreatitis to physiologic autophagy triggered b...

Objectives: Cigarette smoking is a major risk factor for pancreatic cancer (PaCa). However, the mechanisms of smoking-induced PaCa remain unknown. Here we investigated the effect of smoking compounds on cell death pathways in pancreatic ductal cells, precursors of PaCa. Methods: Human pancreatic ductal cells (HPDE6-c7) were cultured with cigaret...

Background & aims: Opening of the mitochondrial permeability transition pore (MPTP) causes loss of the mitochondrial membrane potential (ΔΨm) and, ultimately, adenosine triphosphate depletion and necrosis. Cells deficient in cyclophilin D (CypD), a component of the MPTP, are resistant to MPTP opening, loss of ΔΨm, and necrosis. Alcohol abuse is a...

Acute pancreatitis is an inflammatory disease of exocrine pancreas that carries considerable morbidity and mortality; its pathophysiology remains poorly understood. This review summarizes recent findings obtained in experimental models and genetically altered mice, which reveal that autophagy, the principal cellular degradative pathway, is impaired...

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive disease. The prognosis is poor; less than 5% of those diagnosed are still alive five years after diagnosis, and complete remission is still rare. Tobacco smoking is a major risk factor of pancreatic cancer. However, the mechanism(s) through which it causes the disease remains unknown. Accumul...

Proceedings: AACR 103rd Annual Meeting 2012‐‐ Mar 31‐Apr 4, 2012; Chicago, IL Background & Aims: Cigarette smoking is a major risk factor for pancreatic cancer. However, the molecular signaling mechanisms through which smoking promotes pancreatic cancer remain unknown. Experimental models of smoking-induced pancreatic cancer have not been develope...

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring au...

Inflammation and acinar cell necrosis are two major pathological responses of acute pancreatitis, a serious disorder with no current therapies directed to its molecular pathogenesis. Serine/threonine protein kinase D family, which includes PKD/PKD1, PKD2, and PKD3, has been increasingly implicated in the regulation of multiple physiological and pat...

Recent findings from our group, obtained on experimental in vivo and ex vivo models of pancreatitis, reveal that this disease causes a profound dysfunction of key cellular organelles, lysosomes and mitochondria. We found that autophagy, the main cellular degradative, lysosome-driven process, is activated but also impaired in acute pancreatitis beca...

There are approximately 277,000 new cases of pancreatic cancer and 266,000 deaths from pancreatic cancer annually, indicating a mortality rate of 96% of the cases diagnosed. Because of the ineffectiveness of therapies, a major emphasis needs to be placed on prevention. This paper reviews the epidemiology and risk factors for pancreatic cancer, and...

The kinase Akt mediates resistance of pancreatic cancer (PaCa) cells to death and is constitutively active (phosphorylated) in cancer cells. Whereas the kinases that activate Akt are well characterized, less is known about phosphatases that dephosporylate and thereby inactivate it. We investigated regulation of Akt activity and cell death by the ph...

Significance: Acute pancreatitis is an inflammatory disease of exocrine pancreas that carries considerable morbidity and mortality; its pathophysiology remains poorly understood. During the past decade, new insights have been gained into signaling pathways and molecules that mediate the inflammatory response of pancreatitis and death of acinar cel...

Scutellaria baicalensis (SB) and SB-derived polyphenols possess anti-proliferative activities in several cancers, including pancreatic cancer (PaCa). However, the precise molecular mechanisms have not been fully defined. SB extract and SB-derived polyphenols (wogonin, baicalin, and baicalein) were used to determine their anti-proliferative mechanis...

Background & Aims: Autophagy plays a dual role in cancer development. Recent data indicate that autophagy is oncosuppressive and impaired autophagy is emerging as the critical pro-onco “hallmark” of cancer cells. On the other hand, autophagy is a prosurvival mechanism, and its inhibition may increase cancer cell death and thus be beneficial. Beclin...

Aims: Large doses of intraperitoneally injected basic amino acids, L-arginine, or L-ornithine, induce acute pancreatitis in rodents, although the mechanisms mediating pancreatic toxicity remain unknown. Another basic amino acid, L-lysine, was also shown to cause pancreatic acinar cell injury. The aim of the study was to get insight into the mechan...

Acute pancreatitis is an inflammatory disorder of exocrine pancreas, which carries considerable morbidity and mortality; its pathophysiology remains elusive.1 During the past decade, significant progress has been achieved in our understanding of the inflammatory response in pancreatitis.1 Much less is known about the mechanisms mediating another ke...

Alcohol abuse is one of the most common causes of pancreatitis. The risk of developing alcohol-induced pancreatitis is related to the amount and duration of drinking. However, only a small portion of heavy drinkers develop disease, indicating that other factors (genetic, environmental, or dietary) contribute to disease initiation. Epidemiologic stu...

Proceedings: AACR 101st Annual Meeting 2010‐‐ Apr 17‐21, 2010; Washington, DC Background & Aims: Akt kinase is a major prosurvival factor in pancreatic cancer (PaCa). Akt activity is regulated by its phosphorylation, which is controlled by kinases and phosphatases. Whereas kinases involved in Akt phosphorylation are well characterized, little is k...