Biomedical subjects

Deepak Bhole

Publications and source records attributed to Deepak Bhole.

5 recordsLinked to original sources

PubMed2006

Gene expression analysis of B-lymphoma cells resistant and sensitive to bortezomib.

The proteasome inhibitor bortezomib has shown impressive clinical activity alone and in combination with conventional and other novel agents for the treatment of multiple myeloma (MM). Although bortezomib is known to be a selective proteasome inhibitor, the downstream mechanisms of cytotoxicity and drug resistance are poorly understood. However, resistance to bortezomib as a single agent develops in the majority of patients, and activity in other malignancies has been less impressive. To elucidate mechanisms of bortezomib resistance, we compared differential gene expression profiles of bortezomib-resistant SUDHL-4 and bortezomib-sensitive SUDHL-6 diffuse large B-cell lymphoma lines in response to bortezomib. At concentrations that effectively inhibited proteasome activity, bortezomib induced apoptosis in SUDHL-6 cells, but not in SUDHL-4 cells. We showed that overexpression of activating transcription factor 3 (ATF3), ATF4, ATF5, c-Jun, JunD and caspase-3 is associated with sensitivity to bortezomib-induced apoptosis, whereas overexpression of heat shock protein (HSP)27, HSP70, HSP90 and T-cell factor 4 is associated with bortezomib resistance.

Antineoplastic Agents↗

PubMed2004

Molecular basis for complement component 6 (C6) deficiency in rats and mice.

Complement component C6 is a part of the lytic membrane attack complex formed during complement activation. Animal modeling to define the role of C5a vs. C5b-9 in human disease has used rodents deficient in C6, yet the molecular basis for the deficiencies has not been ascertained. Oligonucleotides derived from a 493 bp EST sequence of the rat C6 gene were used to isolate full-length transcripts of rat C6 mRNA. Sequence analysis confirmed that the derived amino acid sequence for rat C6 is highly homologous to human and mouse. We identified a 31 bp deletion in exon 10 of the C6 gene that leads to C6 deficiency in a strain of PVG rats (PVG/c-) and developed a PCR-based genotyping test. In addition, we identified four point mutations in the mouse C6 gene that may result in C6 deficiency observed in the Peru-Coppock mouse strain. A serendipitous finding from this study was a coagulation defect in the C6 deficient mice and rats. C6 deficient mice or rats demonstrated prolonged tail bleeding times that was reversed by treatment with purified rat C6 protein. Further, adenosine diphosphate induced platelet aggregation were markedly reduced in C6 deficient rats. The molecular basis for these coagulations defects is unknown at present.

Amino Acid Sequence↗

PubMed2004

Doxycycline-regulated over-expression of hsp22 has negative effects on stress resistance and life span in adult Drosophila melanogaster.

Drosophila hsp22 is a member of the small heat shock proteins family (shsps). The hsp22 is expressed in a tissue-general pattern in response to heat stress and during normal aging, and localizes to the mitochondrial matrix, however, its exact function and targets are unknown. Hsp22 was found to be rapidly induced in response to oxidative stress, indicating that hsp22 is also an oxidative stress response gene. To assay for effects of hsp22, a ubiquitous pattern of hsp22 gene expression was generated in young flies using the "tet-on" doxycycline-regulated promoter system. The hsp22 over-expression made flies more sensitive to heat and oxidative stress, while resistance to coumarin poisoning was not affected. Life span was also reduced, particularly at higher culture temperatures. Members of other hsp families have been shown to feedback-inhibit their own expression by interacting with the heat shock transcription factor (HSF) and preventing binding to the HSEs. Induction of hsp22:lacZ and hsp70:lacZ reporter transgenes in response to acute stress was normal in the presence of hsp22 protein over-expression and in old flies, indicating that the negative effects of hsp22 are downstream of the HSF/HSE pathway and the transcriptional heat shock response. The data demonstrate a specific over-expression phenotype for hsp22 and suggest that hsp22 interacts with heat and oxidative stress resistance pathways.

Aging↗

PubMed2003

A search for doxycycline-dependent mutations that increase Drosophila melanogaster life span identifies the VhaSFD, Sugar baby, filamin, fwd and Cctl genes.

BACKGROUND: A P-type transposable element called PdL has been engineered with a doxycycline-inducible promoter directed out through the 3' end of the element. Insertion of PdL near the 5' end of a gene often yields doxycycline-dependent overexpression of that gene and a mutant phenotype. This functional genomics strategy allows for efficient screening of large numbers of genes for overexpression phenotypes. RESULTS: PdL was mobilized to around 10,000 new locations in the Drosophila melanogaster genome and used to search for genes that would extend life span when overexpressed. Six lines were identified in which there was a 5-17% increase in life span in the presence of doxyxcycline. The mutations were molecularly characterized and in each case a gene was found to be overexpressed using northern blots. Two genes did not have previously known phenotypes and are implicated in membrane transport: VhaSFD encodes a regulatory subunit of the vacuolar ATPase proton pump (H+-ATPase), whereas Sugar baby (Sug) is related to a maltose permease from Bacillus. Three PdL mutations identified previously characterized genes: filamin encodes the homolog of an actin-polymerizing protein that interacts with presenilins. four wheel drive (fwd) encodes a phosphatidylinositol-4-kinase (PI 4-kinase) and CTP:phosphocholine cytidylyltransferase-l (Cctl) encodes the rate-limiting enzyme in phosphatidylcholine synthesis. Finally, an apparently novel gene (Red herring, Rdh) was found in the first intron of the encore gene. CONCLUSIONS: Screening for conditional mutations that increase Drosophila life span has identified genes implicated in membrane transport, phospholipid metabolism and signaling, and actin cytoskeleton organization.

1-Phosphatidylinositol 4-Kinase↗

PubMed2003

Therapeutic potential of targeting the complement cascade in critical care medicine.

Caring for the critical care patient involves many different areas of clinical expertise and serves a diverse patient population. Novel therapeutics for the critically ill must be approached with caution, because the underlying molecular mechanisms of the disease process for several commonly seen types of patients (i.e., sepsis, shock, ischemia/reperfusion) are not fully understood. A potentially new and advancing area of therapeutics that may hold promise for the critically ill is inhibition of the complement system. Various novel complement inhibitors are being developed and several are in clinical trials. The advancement of this novel area of therapeutics may one day aid the clinician by providing several different complement inhibitors/antagonists for controlling complement activation or its biologically active mediators.

Complement Inactivator Proteins↗