Priit Kogerman - Academia.edu (original) (raw)

Papers by Priit Kogerman

Journal of Controlled Release, 2005

Cell-penetrating peptides (CPPs) are novel high-capacity delivery vectors for different bioactive... more Cell-penetrating peptides (CPPs) are novel high-capacity delivery vectors for different bioactive cargoes. We have evaluated the CPP transportan 10 (TP10) as a delivery vector in different in vitro plasmid delivery assays. Tested methods include: TP10 crosslinked to a plasmid via a peptide nucleic acid (PNA) oligomer, TP10 conjugation with polyethyleneimine (PEI), and addition of unconjugated TP10 to standard PEI transfection assay. We found that without additional DNA condensing agents, TP10 has poor transfection abilities. However, the presence of TP10 increases the transfection efficiency several folds compared to PEI alone. At as low concentrations as 0.6 nM, TP10-PNA constructs were found to enhance plasmid delivery up to 3.7-fold in Neuro-2a cells. Interestingly, the transfection efficiency was most significant at low PEI concentrations, allowing reduced PEI concentration without loss of gene delivery. No increase in cytotoxicity due to TP10 was observed and the uptake mechanism was determined to be endocytosis, as previously reported for PEI mediated transfection. In conclusion, TP10 can enhance PEI mediated transfection at relatively low concentrations and may help to develop future gene delivery systems with reduced toxicity.

Plant Signaling & Behavior, 2008

International Journal of Immunotherapy, 1997

Use of a molecule comprising a variant that does not bind to the HA binding domain hyaluronan CD4... more Use of a molecule comprising a variant that does not bind to the HA binding domain hyaluronan CD44 (CD44-HABD), as well as analog, recombinant and mutated variants or fragments thereof, for the manufacture of a medicament for treating conditions related to inhibition of angiogenesis or endothelial cell proliferation, such as ocular diseases causing blindness or visual disturbances, such as macular degeneration, diabetic retinopathy and states of retinal hypoxia; states of chronic inflammation, such as rheumatoid arthritis, psoriasis, atherosclerosis, restenosis; as well as in cancer growth and metastasis, as well as all forms of cancer and tumors, such as breast, prostate, colon, lung, skin, liver, brain, ovary, testis, skeleton, epithelium, endothelium, pancreas , kidney, muscle, adrenal gland, intestines, endocrine glands, oral cavity, head and neck and other solid tissue origin, or any form of leukemia, as well as hemangiomas, wherein the binding domain hyaluronan CD44 has homolo...

La presente invention concerne un procede permettant de predire ou concevoir, detecter et/ou veri... more La presente invention concerne un procede permettant de predire ou concevoir, detecter et/ou verifier un nouveau peptide penetrant dans les cellules (CPP) et un procede permettant d'utiliser ce nouveau peptide CPP et/ou un nouvel usage d'un CPP connu pour une meilleure absorption cellulaire d'un effecteur cellulaire couple a ce peptide CPP. La presente invention concerne egalement un procede de prediction ou de conception, detection et/ou verification d'un nouveau peptide penetrant dans les cellules (CPP) qui imitent l'activite de l'effecteur cellulaire et/ou inhibent l'activite de l'effecteur cellulaire. La presente invention concerne enfin l'utilisation de ce CPP pour traiter et/ou prevenir un etat medical et l'utilisation de ce CPP pour la production d'une composition pharmaceutique en vue de traiter un etat medical.

Proceedings of the National Academy of Sciences, 1999

Truncation mutations of the GLI3 zinc finger transcription factor can cause Greig cephalopolysynd... more Truncation mutations of the GLI3 zinc finger transcription factor can cause Greig cephalopolysyndactyly syndrome (GCPS), Pallister–Hall syndrome (PHS), and postaxial polydactyly type A (PAP-A). GLI3 is homologous to Drosophila Cubitus interruptus (Ci), which regulates the patched ( ptc ), gooseberry ( gsb ), and decapentaplegic ( dpp ) genes. Ci is sequestered in the cytoplasm and is subject to posttranslational processing whereby the full-length transcriptional activator form (Ci 155 ) can be cleaved to a repressor form (Ci 75 ). Under hedgehog signaling, the Ci 155 form translocates to the nucleus whereas in the absence of hedgehog, the Ci 75 form translocates to the nucleus. Based on the correlation of GLI3 truncation mutations and the human phenotypes, we hypothesized that GLI3 shows transcriptional activation or repression activity and subcellular localization similar to Ci. Here we show that full-length GLI3 localizes to the cytoplasm and activates PTCH1 expression, which is s...

Hybridoma, 2008

Gli3 is a key regulator of development, controlling multiple patterning steps. Here we report the... more Gli3 is a key regulator of development, controlling multiple patterning steps. Here we report the generation of a scFv antibody specific to the repressor domain of human Gli3. We show that this scFv retains the binding capacity of its parent anti-Gli3 monoclonal antibody derived from hybridoma clone 5E1. When expressed in mammalian cells, the anti-Gli3 scFv co-localizes with intracellular Gli3. Immunocytochemical staining of the intrabody in Gli3-positive TM4 cells shows a distinct perinuclear cytoplasmic localization. Such a scFv constitutes a useful tool for studying transcriptional regulation of the hedgehog pathway in mammals and offers a starting point for developing novel Gli-related therapeutic intrabodies.

Journal of Biological Chemistry, 2002

Oncogene, 1997

Human CD44 standard isoform cDNA (hCD44s) was transfected into sis-transformed Balb/c 3T3 cells a... more Human CD44 standard isoform cDNA (hCD44s) was transfected into sis-transformed Balb/c 3T3 cells and into ras-revertant IIIA4 cells (both tumorigenic but nonmetastatic). Transfectants were injected subcutaneously into athymic nude mice to elucidate the functional role of hCD44s over-expression in progression and metastasis. The transfectants (but not parental cells) were capable of lung micrometastasis and of binding exogenously-added hyaluronan. hCD44s protein expression was conserved in lung micrometastases suggesting that it may have been necessary for their formation. In contrast, no hCD44s protein was detected in large subcutaneous (s.c.) tumors but normal levels of murine CD44 were detected. A second round of tumor development, using these two tumor cell classes, demonstrated that hCD44s-nonexpressing s.c. tumor cells re-expressed it in lung micrometastases. Conversely, hCD44s-expressing lung micrometastatic cells, when injected into a second group of mice, downregulated hCD44s expression in order to grow sizable s.c. tumors. S.c. tumor cells still contained the hCD44s gene but its expression was inhibited by epigenetic mechanisms, one of which was shown to be methylation of the hCD44s gene. These studies demonstrate (a) opposing selective pressures on CD44s over-expression for s.c. tumor growth and for metastatic spread to the lung and (b) further credence for the signi®cance of CD44 for metastatic spread of ®brosarcomas. Therefore, CD44s may be a critical component of the metastatic phenotype induced by speci®c oncogenes.

<b>Copyright information:</b>Taken from "A potential role of alternative splicin... more <b>Copyright information:</b>Taken from "A potential role of alternative splicing in the regulation of the transcriptional activity of human GLI2 in gonadal tissues"BMC Molecular Biology 2006;7():13-13.Published online 23 Mar 2006PMCID:PMC1435915.erence accession numbers shown on the right). Exons (numbered) and introns are shown by vertical and horizontal lines, respectively. Alternative 5' noncoding exons in human gene are designated as 1a and 1b. Asterisks indicate exons 3–6, predicted from the comparison of mouse and human Gli2 mRNAs and genomic structures, identified by Roessler et al. [33].

CD44 is a cell surface glycoprotein that functions as hyaluronan receptor. Mouse and human serum ... more CD44 is a cell surface glycoprotein that functions as hyaluronan receptor. Mouse and human serum contain substantial amounts of soluble CD44, generated either by shedding or alternative splicing. During inflammation and in cancer patients serum levels of soluble CD44 are significantly increased. Experimentally, soluble CD44 overexpression blocks cancer cell adhesion to HA. We have previously found that recombinant CD44 hyaluronan binding domain (CD44HABD) and its nonHA-binding mutant inhibited tumor xenograft growth, angiogenesis, and endothelial cell proliferation. These data suggested an additional target other than HA for CD44HABD. By using non-HA-binding CD44HABD Arg41Ala, Arg78Ser, and Tyr79Ser-triple mutant (CD443MUT) we have identified intermediate filament protein vimentin as a novel interaction partner of CD44. We found that vimentin is expressed on the cell surface of human umbilical vein endothelial cells (HUVEC). Endogenous CD44 and vimentin coprecipitate from HUVECs, an...

The segment polarity gene Fused (Fu) encodes a putative serine–threonine kinase Fu, which has bee... more The segment polarity gene Fused (Fu) encodes a putative serine–threonine kinase Fu, which has been shown to play a key role in the Hedgehog signaling pathway of Drosophila. Human FU (hFU) has been shown to enhance the activity of Gli transcription factors, targets of the signaling pathway. However, Fu mice do not show aberrant embryonic development indicating that mouse Fu (mFu) is dispensable for Hedgehog signaling until birth. In order to investigate if there are important differences between hFU and mFu, we cloned the cDNA, analyzed expression and tested the ability of mFu to regulate Gli proteins. Of the tested tissues only brain and testis showed significant expression. However, in transient overexpression analyses mFu was able to enhance Gli induced transcription in a manner similar to hFU. Thus, we turned to RNAi in order to test if mFu would be important for Hedgehog signaling after all. In one cell line with reduced mFu expression the Hedgehog signaling was severely hampere...

Journal of Controlled Release, 2005

Cell-penetrating peptides (CPPs) are novel high-capacity delivery vectors for different bioactive... more Cell-penetrating peptides (CPPs) are novel high-capacity delivery vectors for different bioactive cargoes. We have evaluated the CPP transportan 10 (TP10) as a delivery vector in different in vitro plasmid delivery assays. Tested methods include: TP10 crosslinked to a plasmid via a peptide nucleic acid (PNA) oligomer, TP10 conjugation with polyethyleneimine (PEI), and addition of unconjugated TP10 to standard PEI transfection assay. We found that without additional DNA condensing agents, TP10 has poor transfection abilities. However, the presence of TP10 increases the transfection efficiency several folds compared to PEI alone. At as low concentrations as 0.6 nM, TP10-PNA constructs were found to enhance plasmid delivery up to 3.7-fold in Neuro-2a cells. Interestingly, the transfection efficiency was most significant at low PEI concentrations, allowing reduced PEI concentration without loss of gene delivery. No increase in cytotoxicity due to TP10 was observed and the uptake mechanism was determined to be endocytosis, as previously reported for PEI mediated transfection. In conclusion, TP10 can enhance PEI mediated transfection at relatively low concentrations and may help to develop future gene delivery systems with reduced toxicity.

Plant Signaling & Behavior, 2008

International Journal of Immunotherapy, 1997

Use of a molecule comprising a variant that does not bind to the HA binding domain hyaluronan CD4... more Use of a molecule comprising a variant that does not bind to the HA binding domain hyaluronan CD44 (CD44-HABD), as well as analog, recombinant and mutated variants or fragments thereof, for the manufacture of a medicament for treating conditions related to inhibition of angiogenesis or endothelial cell proliferation, such as ocular diseases causing blindness or visual disturbances, such as macular degeneration, diabetic retinopathy and states of retinal hypoxia; states of chronic inflammation, such as rheumatoid arthritis, psoriasis, atherosclerosis, restenosis; as well as in cancer growth and metastasis, as well as all forms of cancer and tumors, such as breast, prostate, colon, lung, skin, liver, brain, ovary, testis, skeleton, epithelium, endothelium, pancreas , kidney, muscle, adrenal gland, intestines, endocrine glands, oral cavity, head and neck and other solid tissue origin, or any form of leukemia, as well as hemangiomas, wherein the binding domain hyaluronan CD44 has homolo...

La presente invention concerne un procede permettant de predire ou concevoir, detecter et/ou veri... more La presente invention concerne un procede permettant de predire ou concevoir, detecter et/ou verifier un nouveau peptide penetrant dans les cellules (CPP) et un procede permettant d'utiliser ce nouveau peptide CPP et/ou un nouvel usage d'un CPP connu pour une meilleure absorption cellulaire d'un effecteur cellulaire couple a ce peptide CPP. La presente invention concerne egalement un procede de prediction ou de conception, detection et/ou verification d'un nouveau peptide penetrant dans les cellules (CPP) qui imitent l'activite de l'effecteur cellulaire et/ou inhibent l'activite de l'effecteur cellulaire. La presente invention concerne enfin l'utilisation de ce CPP pour traiter et/ou prevenir un etat medical et l'utilisation de ce CPP pour la production d'une composition pharmaceutique en vue de traiter un etat medical.

Proceedings of the National Academy of Sciences, 1999

Truncation mutations of the GLI3 zinc finger transcription factor can cause Greig cephalopolysynd... more Truncation mutations of the GLI3 zinc finger transcription factor can cause Greig cephalopolysyndactyly syndrome (GCPS), Pallister–Hall syndrome (PHS), and postaxial polydactyly type A (PAP-A). GLI3 is homologous to Drosophila Cubitus interruptus (Ci), which regulates the patched ( ptc ), gooseberry ( gsb ), and decapentaplegic ( dpp ) genes. Ci is sequestered in the cytoplasm and is subject to posttranslational processing whereby the full-length transcriptional activator form (Ci 155 ) can be cleaved to a repressor form (Ci 75 ). Under hedgehog signaling, the Ci 155 form translocates to the nucleus whereas in the absence of hedgehog, the Ci 75 form translocates to the nucleus. Based on the correlation of GLI3 truncation mutations and the human phenotypes, we hypothesized that GLI3 shows transcriptional activation or repression activity and subcellular localization similar to Ci. Here we show that full-length GLI3 localizes to the cytoplasm and activates PTCH1 expression, which is s...

Hybridoma, 2008

Gli3 is a key regulator of development, controlling multiple patterning steps. Here we report the... more Gli3 is a key regulator of development, controlling multiple patterning steps. Here we report the generation of a scFv antibody specific to the repressor domain of human Gli3. We show that this scFv retains the binding capacity of its parent anti-Gli3 monoclonal antibody derived from hybridoma clone 5E1. When expressed in mammalian cells, the anti-Gli3 scFv co-localizes with intracellular Gli3. Immunocytochemical staining of the intrabody in Gli3-positive TM4 cells shows a distinct perinuclear cytoplasmic localization. Such a scFv constitutes a useful tool for studying transcriptional regulation of the hedgehog pathway in mammals and offers a starting point for developing novel Gli-related therapeutic intrabodies.

Journal of Biological Chemistry, 2002

Oncogene, 1997

Human CD44 standard isoform cDNA (hCD44s) was transfected into sis-transformed Balb/c 3T3 cells a... more Human CD44 standard isoform cDNA (hCD44s) was transfected into sis-transformed Balb/c 3T3 cells and into ras-revertant IIIA4 cells (both tumorigenic but nonmetastatic). Transfectants were injected subcutaneously into athymic nude mice to elucidate the functional role of hCD44s over-expression in progression and metastasis. The transfectants (but not parental cells) were capable of lung micrometastasis and of binding exogenously-added hyaluronan. hCD44s protein expression was conserved in lung micrometastases suggesting that it may have been necessary for their formation. In contrast, no hCD44s protein was detected in large subcutaneous (s.c.) tumors but normal levels of murine CD44 were detected. A second round of tumor development, using these two tumor cell classes, demonstrated that hCD44s-nonexpressing s.c. tumor cells re-expressed it in lung micrometastases. Conversely, hCD44s-expressing lung micrometastatic cells, when injected into a second group of mice, downregulated hCD44s expression in order to grow sizable s.c. tumors. S.c. tumor cells still contained the hCD44s gene but its expression was inhibited by epigenetic mechanisms, one of which was shown to be methylation of the hCD44s gene. These studies demonstrate (a) opposing selective pressures on CD44s over-expression for s.c. tumor growth and for metastatic spread to the lung and (b) further credence for the signi®cance of CD44 for metastatic spread of ®brosarcomas. Therefore, CD44s may be a critical component of the metastatic phenotype induced by speci®c oncogenes.

<b>Copyright information:</b>Taken from "A potential role of alternative splicin... more <b>Copyright information:</b>Taken from "A potential role of alternative splicing in the regulation of the transcriptional activity of human GLI2 in gonadal tissues"BMC Molecular Biology 2006;7():13-13.Published online 23 Mar 2006PMCID:PMC1435915.erence accession numbers shown on the right). Exons (numbered) and introns are shown by vertical and horizontal lines, respectively. Alternative 5' noncoding exons in human gene are designated as 1a and 1b. Asterisks indicate exons 3–6, predicted from the comparison of mouse and human Gli2 mRNAs and genomic structures, identified by Roessler et al. [33].

CD44 is a cell surface glycoprotein that functions as hyaluronan receptor. Mouse and human serum ... more CD44 is a cell surface glycoprotein that functions as hyaluronan receptor. Mouse and human serum contain substantial amounts of soluble CD44, generated either by shedding or alternative splicing. During inflammation and in cancer patients serum levels of soluble CD44 are significantly increased. Experimentally, soluble CD44 overexpression blocks cancer cell adhesion to HA. We have previously found that recombinant CD44 hyaluronan binding domain (CD44HABD) and its nonHA-binding mutant inhibited tumor xenograft growth, angiogenesis, and endothelial cell proliferation. These data suggested an additional target other than HA for CD44HABD. By using non-HA-binding CD44HABD Arg41Ala, Arg78Ser, and Tyr79Ser-triple mutant (CD443MUT) we have identified intermediate filament protein vimentin as a novel interaction partner of CD44. We found that vimentin is expressed on the cell surface of human umbilical vein endothelial cells (HUVEC). Endogenous CD44 and vimentin coprecipitate from HUVECs, an...

The segment polarity gene Fused (Fu) encodes a putative serine–threonine kinase Fu, which has bee... more The segment polarity gene Fused (Fu) encodes a putative serine–threonine kinase Fu, which has been shown to play a key role in the Hedgehog signaling pathway of Drosophila. Human FU (hFU) has been shown to enhance the activity of Gli transcription factors, targets of the signaling pathway. However, Fu mice do not show aberrant embryonic development indicating that mouse Fu (mFu) is dispensable for Hedgehog signaling until birth. In order to investigate if there are important differences between hFU and mFu, we cloned the cDNA, analyzed expression and tested the ability of mFu to regulate Gli proteins. Of the tested tissues only brain and testis showed significant expression. However, in transient overexpression analyses mFu was able to enhance Gli induced transcription in a manner similar to hFU. Thus, we turned to RNAi in order to test if mFu would be important for Hedgehog signaling after all. In one cell line with reduced mFu expression the Hedgehog signaling was severely hampere...