Daniel Emerling - Academia.edu (original) (raw)

Papers by Daniel Emerling

Journal of Biological Chemistry, Jul 1, 1994

Previously, we characterized the prostaglandin (PG) F2 alpha receptor linked to phospholipase C a... more Previously, we characterized the prostaglandin (PG) F2 alpha receptor linked to phospholipase C activation and DNA synthesis in NIH-3T3 cells (Nakao, A., Watanabe, T., Taniguchi, S., Nakamura, M., Honda, Z-I., Shimizu, T., and Kurokawa, K. (1993) J. Cell. Physiol. 155, 257-264). To elucidate intracellular events evoked via this receptor, we examined changes caused by PGF2 alpha stimulation in the phosphotyrosine composition of cellular proteins. The addition of PGF2 alpha to cells in quiescent culture rapidly increased the levels of phosphotyrosine in cellular proteins with Mr values of 70,000 (pp70), 85,000 (pp85), 92,000 (pp92), 100,000 (pp100), and 125,000 (pp125); the latter was immunologically identified as p125 focal adhesion kinase. The PGF2 alpha-induced changes in the level of intracellular Ca2+ ([Ca2+]i) elevation, formation of inositol phosphates, and [3H]thymidine incorporation followed a similar dose dependence as PGF2 alpha-induced tyrosine phosphorylation. This tyrosine phosphorylation was independent of extracellular Ca2+, while a [Ca2+]i chelator, 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (50 microM), completely inhibited the PGF2 alpha-induced elevation of [Ca2+]i, tyrosine phosphorylation, and [3H]thymidine incorporation. Ionomycin (0.1 microM), which induced [Ca2+]i elevation without formation of inositol phosphates, mimicked the PGF2 alpha-induced tyrosine phosphorylation. 12-O-Tetradecanoylphorbol-13-acetate (TPA) also induced [3H]thymidine incorporation in a dose-dependent manner but had no significant effect on tyrosine phosphorylation. The PGF2 alpha-induced tyrosine phosphorylation could be observed even in the cells pretreated with TPA (5 microM, 24 h). PGF2 alpha exhibited an additive effect on TPA-induced [3H]thymidine incorporation but had no effect on the 32P-phosphorylation of a known 80-kDa protein kinase (PK) C substrate. Both staurosporine and H-7 inhibited the PGF2 alpha-induced increase in [3H]thymidine incorporation and tyrosine phosphorylation in a similar dose-dependent manner whether or not cells were pretreated with TPA (5 microM, 24 h). However, W-7 and KN-62 had no effect on these cellular responses even at the concentration for the almost complete inhibition of Ca2+/calmodulin-dependent PKs (20 microM). These results, taken together, indicate that PGF2 alpha receptor-mediated tyrosine phosphorylation is evoked by a [Ca2+]i-dependent mechanism that is sensitive to staurosporine and H-7 but which is independent of PKC or Ca2+/calmodulin PKs. Finally, the data suggest that this PGF2 alpha-induced signaling pathway is linked to the proliferation of cells.

Journal of Clinical Oncology, May 20, 2016

e23113Background: Analyzing anti-cancer immune responses gives insights into the mechanisms that ... more e23113Background: Analyzing anti-cancer immune responses gives insights into the mechanisms that underlie successful cancer immunotherapies, and may find potentially useful antibodies. Methods: To ...

Nature Communications, Jul 28, 2023

Research Square (Research Square), Apr 4, 2023

Over 80% of malaria-attributable deaths are in children under ve. However, the only malaria vacci... more Over 80% of malaria-attributable deaths are in children under ve. However, the only malaria vaccine recommended by the World Health Organization (WHO) for paediatric use, Mosquirix™, has limited e cacy. Complementary strategies, like monoclonal antibodies (mAbs), will be required to eradicate malaria. To discover new anti-malaria mAbs, we evaluated >28,000 antibody sequences from circulating B cells obtained from 45 Mosquirix™ vaccinees and selected 369 for testing. Many antibodies bound the circumsporozoite protein (CSP), a main surface protein on malaria and the malaria antigen in Mosquirix™, and several were exceptionally protective in mouse models of malaria. Through this work, we identi ed surprising correlations that suggest certain CSP sequences in Mosquirix™ may induce immunodominant antibody responses that dilute protective immunity. Further, we selected the antibodies most protective in preclinical mouse models and engineered them for improved manufacturability and developability to meet WHO guidelines. An optimised clinical candidate, MAM01, suitable for paediatric populations living in low-to-middle-income countries, was selected for clinical development.

Arthritis & rheumatology, Mar 7, 2023

Journal of Clinical Oncology, May 20, 2020

TPS3168 Background: ATRC-101 is a fully human, engineered IgG1 version of an antibody discovered ... more TPS3168 Background: ATRC-101 is a fully human, engineered IgG1 version of an antibody discovered through a a target-agnostic screen to identify patient-derived antibodies that bind selectively to public tumor antigens. The parental antibody was identified from B cells in the active immune response of a patient receiving checkpoint therapy for Stage IV non-small cell lung cancer (NSCLC). A fluorescently conjugated version of ATRC-101 binds selectively to human tumor specimens including a majority of NSCLC, acral melanoma, breast, colorectal, and ovarian cancer samples. No reactivity of toxicological significance is found across a wide range of normal human tissues. ATRC-101 displays dose-dependent, single-agent activity in syngeneic mouse tumor models, including the EMT6 breast cancer model, which displays a T cell-excluded microenvironment often observed in human tumors, and in which checkpoint inhibitors targeting the PD-1 axis exhibit limited activity. Dosing with ATRC-101 in the EMT6 model causes marked changes in the tumor microenvironment, including a shift from the M2 to the M1 macrophage phenotype and infiltration of T cells. ATRC-101 does not appear to act via NK cell-driven ADCC; instead, activity in vivo is dependent both on Fc region interactions with Fc receptors, likely on myeloid rather than lymphoid cells, and on the presence of CD8+ T cells. ATRC-101 binds to a target that is a ribonucleoprotein (RNP) complex containing polyadenylate-binding protein 1 (PABP-1) bound to poly(A)RNA. Whereas both PABP-1 and poly(A)RNA are ubiquitously expressed at high levels in normal tissues and have been localized intracellularly, the ATRC-101 target is detected extracellularly on tumor cells grown in vivo. The basis for the tumor-selectivity of ATRC-101 as well as the extracellular localization of the target is under investigation. Ascending doses of ATRC-101 were well tolerated in multiple non-clinical safety studies. Methods: ATRC-101-A01 is an open-label, 3+3, Phase 1b safety study in patients with acral melanoma, NSCLC, breast, ovarian, and colorectal cancers. Participants are accruing in the first dose cohort. ATRC-101 is administered every 21 days up to 24 months or until disease progression. The primary objective of the trial is to determine the safety and tolerability of ATRC-101. Secondary objectives are to characterize the pharmacokinetic profiles of ATRC-101 and to assess antitumor activity as determined by RECIST 1.1 and lymphocytic infiltration in the tumor microenvironment. Clinical trial information: NCT04244552 .

Frontiers in Immunology, Oct 25, 2019

Mutants of a catalytically inactive variant of Proteinase 3 (PR3)-iPR3-Val 103 possessing a Ser19... more Mutants of a catalytically inactive variant of Proteinase 3 (PR3)-iPR3-Val 103 possessing a Ser195Ala mutation relative to wild-type PR3-Val 103-offer insights into how autoantigen PR3 interacts with antineutrophil cytoplasmic antibodies (ANCAs) in granulomatosis with polyangiitis (GPA) and whether such interactions can be interrupted. Here we report that iHm5-Val 103 , a triple mutant of iPR3-Val 103 , bound a monoclonal antibody (moANCA518) from a GPA patient on an epitope remote from the mutation sites, whereas the corresponding epitope of iPR3-Val 103 was latent to moANCA518. Simulated B-factor analysis revealed that the binding of moANCA518 to iHm5-Val 103 was due to increased main-chain flexibility of the latent epitope caused by remote mutations, suggesting rigidification of epitopes with therapeutics to alter pathogenic PR3•ANCA interactions as new GPA treatments.

PLOS Pathogens, Mar 28, 2022

Potent and durable vaccine responses will be required for control of malaria caused by Plasmodium... more Potent and durable vaccine responses will be required for control of malaria caused by Plasmodium falciparum (Pf). RTS,S/AS01 is the first, and to date, the only vaccine that has demonstrated significant reduction of clinical and severe malaria in endemic cohorts in Phase 3 trials. Although the vaccine is protective, efficacy declines over time with kinetics paralleling the decline in antibody responses to the Pf circumsporozoite protein (PfCSP). Although most attention has focused on antibodies to repeat motifs on PfCSP, antibodies to other regions may play a role in protection. Here, we expressed and characterized seven monoclonal antibodies to the C-terminal domain of CSP (ctCSP) from volunteers immunized with RTS,S/AS01. Competition and crystal structure studies indicated that the antibodies target two different sites on opposite faces of ctCSP. One site contains a polymorphic region (denoted α-ctCSP) and has been previously characterized, whereas the second is a previously undescribed site on the conserved β-sheet face of the ctCSP (denoted β-ctCSP). Antibodies to the β-ctCSP site exhibited broad reactivity with a diverse panel of ctCSP peptides

Journal of Molecular Biology, Feb 1, 2020

This is a PDF file of an article that has undergone enhancements after acceptance, such as the ad... more This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

Regular and Young Investigator Award Abstracts, Nov 1, 2022

Journal for ImmunoTherapy of Cancer, Nov 1, 2020

Journal of Clinical Oncology, May 20, 2018

e15161Background: Analyzing anti-cancer immune responses can offer insights into the mechanisms t... more e15161Background: Analyzing anti-cancer immune responses can offer insights into the mechanisms that underlie successful cancer therapies. We studied the B and T cell responses of 11 metastatic bre...

Patient immune response to tumor can drive profound and durable clinical benefit, but the contrib... more Patient immune response to tumor can drive profound and durable clinical benefit, but the contribution of antibodies to this benefit is poorly understood. To further our understanding of the role of the humoral response, we compared antibody sequence repertoires of durable responder and non-responder melanoma patients before and during checkpoint inhibitor treatment. We collected pre- and on-treatment peripheral blood from 26 melanoma patients treated with pembrolizumab (9), ipilimumab (8) or nivolumab+ipilimumab (9). Of the 26 patients, 14 were durable responders (RECIST stable disease, partial response, or complete response for at least 6 months) and 12 were non-responders. We generated natively paired heavy and light chain antibody sequences from individual IgG plasmablasts (CD19+CD20-CD38+CD3-CD14-IgA-IgM-IgD-). Sequences were obtained from a total of 26,725 plasmablasts. These sequences were used to reconstruct lineages (sets of plasmablasts likely derived from a common progenitor B cell) where each lineage shares heavy and light V genes, CDR3 length and 80% sequence similarity in CDR3s. Durable responders exhibited an increase in somatic hypermutation (SHM) after initiation of treatment. In contrast, no significant change in SHM was observed in non-responders. Higher SHM was also observed in on-treatment repertoires of responders compared with those of non-responders suggesting that T cell checkpoint inhibitors promote activation of humoral immunity in clinical responders. Furthermore, persistent antibody lineages (observed both before and during treatment) showed higher SHM than lineages observed at only a single time point. Comparison of lineages between patients identified antibodies with high sequence similarity suggesting these antibodies may have arisen from convergent selection, i.e., different patients raising antibodies against shared or similar epitopes. These putative convergent antibodies were enriched in IgG2. IgG2 usage was also higher overall in durable responders than non-responders, even in samples taken prior to treatment. The higher levels of IgG2 observed in responders and in the potentially convergent sets of antibodies suggest that IgG2 antibodies may play a role in effective anti-tumor responses. In conclusion, our analysis of plasmablast repertoires from melanoma patients suggests that treatment with checkpoint inhibitors promotes SHM in durable responders, and that responder plasmablasts are enriched in IgG2. Moreover, patients make potentially convergent antibodies that are also enriched for the IgG2 subclass. Our observations demonstrate that the humoral immune response is remodeled in patients with anti-tumor responses and support a role for the humoral arm of the immune response in driving clinical benefit in patients treated with checkpoint inhibitors. Citation Format: Ngan Nguyen, Alusha Mamchak, Mariano Severgnini, Kevin S. Williamson, Xinqi Wu, Elliott F. Drabek, Michael Manos, Xiaomu Chen, Xiaobin Tang, Zoe Amiri, Chantia Carroll, Yvonne Leung, Dongkyoon Kim, Wayne Volkmuth, Norman Greenberg, Daniel Emerling, William H. Robinson, Guy Cavet, F. Stephen Hodi. Increased somatic hypermutation in the immunoglobulin sequences of melanoma patients who have durable response to checkpoint inhibitor therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 615.

Proceedings of the National Academy of Sciences of the United States of America, Jun 11, 2018

E10438-E10445). The authors note that they inadvertently omitted references that reported on comp... more E10438-E10445). The authors note that they inadvertently omitted references that reported on computational models of helical structures for the NANP repeats of circumsporozoite protein (CSP) (1-3), one of which is a recent publication that used a helical model of (NANP)6 to explain the multivalent CSP binding of an anti-NANP antibody (3). The model described in Oyen et al., however, differs extensively from these previous models in that the NANP repeats adopt a wide, elongated spiral, compared for example to the tightly packed helix as described in ref. 3. The full references 1-3 appear below. The references should have been cited in an additional sentence, which should have appeared on page E10443, left column, first paragraph, to precede the sentence that begins on line 3 with "Many questions remain.. . ." The additional sentence should have appeared as follows: "Helical models have been predicted previously for the NANP repeats (1-3), but these differ substantially from the spiral conformation presented here." The authors also note that Fig. S7 appeared incorrectly. The corrected Supporting figure and its legend appear below. The SI has been corrected online.

SummaryThe generation of high-quality antibody responses to PfCSP, the primary surface antigen of... more SummaryThe generation of high-quality antibody responses to PfCSP, the primary surface antigen ofPlasmodium falciparumsporozoites, is paramount to the development of an effective malaria vaccine. Here we present an in-depth structural and functional analysis of a panel of potent antibodies encoded by theIGHV3-33germline gene, which is among the most prevalent and potent antibody families induced in the anti-CSP immune response and targets the NANP repeat region. Cryo-EM reveals a remarkable spectrum of helical Fab-CSP structures stabilized by homotypic interactions between tightly packed Fabs, many of which correlate with somatic hypermutation. We demonstrate a key role of these mutated homotypic contacts for high avidity binding to CSP and in protection fromP. falciparummalaria infection. These data emphasize the importance of anti-homotypic affinity maturation in the frequent selection ofIGHV3-33antibodies, advance our understanding of the mechanism(s) of antibody-mediated protect...

Cancer Research, 2021

Background: ATRC-101 is a fully human, engineered IgG1 version of an antibody discovered through ... more Background: ATRC-101 is a fully human, engineered IgG1 version of an antibody discovered through a target-agnostic process designed to identify patient-derived, tumor-targeting antibodies. The parental antibody of ATRC-101 was discovered from a patient with metastatic non-small cell lung cancer (NSCLC) undergoing an active immune response while receiving a checkpoint inhibitor. ATRC-101 binds selectively to human tumor specimens, including a majority of NSCLC, acral melanoma, breast, colorectal, and ovarian cancer samples. The target of ATRC-101 appears to be a tumor-associated ribonucleoprotein complex containing a form of polyadenylate-binding protein 1 (PABP-1) that can be found on the surface of tumor cells. Studies to further elucidate the tumor selectivity of ATRC-101 and the extracellular presentation of the target are ongoing. Preclinical data suggest that ATRC-101 stimulates an adaptive immune response against tumors via the innate immune system. ATRC-101 displays dose-depe...

Journal of Clinical Oncology, 2018

e15015Background: It is unclear exactly how B cells and their antibodies (Abs) influence the anti... more e15015Background: It is unclear exactly how B cells and their antibodies (Abs) influence the anti-cancer immune response. We've proposed that anti-tumor Abs drive or amplify anti-tumor immune effec...

Nature Communications, 2019

Transmission-blocking vaccines have the potential to be key contributors to malaria elimination. ... more Transmission-blocking vaccines have the potential to be key contributors to malaria elimination. Such vaccines elicit antibodies that inhibit parasites during their development in Anopheles mosquitoes, thus breaking the cycle of transmission. To date, characterization of humoral responses to Plasmodium falciparum transmission-blocking vaccine candidate Pfs25 has largely been conducted in pre-clinical models. Here, we present molecular analyses of human antibody responses generated in a clinical trial evaluating Pfs25 vaccination. From a collection of monoclonal antibodies with transmission-blocking activity, we identify the most potent transmission-blocking antibody yet described against Pfs25; 2544. The interactions of 2544 and three other antibodies with Pfs25 are analyzed by crystallography to understand structural requirements for elicitation of human transmission-blocking responses. Our analyses provide insights into Pfs25 immunogenicity and epitope potency, and detail an affin...