scispace - formally typeset
Search or ask a question
Journal Article•DOI•

Activation of Akt is associated with poor prognosis and chemotherapeutic resistance in pediatric B-precursor acute lymphoblastic leukemia.

Naoto Morishita1, Hirokazu Tsukahara1, Kosuke Chayama, Toshiaki Ishida2, Kana Washio1, Takako Miyamura1, Nobuko Yamashita1, Megumi Oda1, Tsuneo Morishima1 •
Okayama University1, Boston Children's Hospital2
15 Jul 2012-Pediatric Blood & Cancer (Pediatr Blood Cancer)-Vol. 59, Iss: 1, pp 83-89
TL;DR: This study was undertaken to explore the clinical relevance and molecular mechanisms underlying the activation of Akt (i.e., phosphorylated Akt, P‐Akt) in pediatric B‐pre ALL.
Abstract: Background Activation of the phosphoinositide 3-kinase (PI3K)/Akt pathway, a pro-survival pathway, plays important roles in tumor cell growth. However, the role of Akt in the pathogenesis of pediatric B-precursor acute lymphoblastic leukemia (B-pre ALL) remains to be clarified. This study was undertaken to explore the clinical relevance and molecular mechanisms underlying the activation of Akt (i.e., phosphorylated Akt, P-Akt) in pediatric B-pre ALL. Procedure We evaluated the activation status of Akt in bone marrow samples from 21 children with newly diagnosed B-pre ALL and correlated the expression level of P-Akt with clinicopathologic and prognostic features. Additionally, we transfected the myristoylated Akt cDNA into the B-pre ALL cell line, Nalm-6, and examined the effect, in vitro, of Akt activation on the response to antitumor drugs. Results P-Akt expression in B-pre ALL blast cells at diagnosis was associated significantly with poor response to induction chemotherapy including prednisolone, dexamethasone, vincristine, and adriamycin in B-pre ALL patients. Both overall survival and relapse-free survival in patients with P-Akt expression were reduced significantly more than in patients without P-Akt expression. Activation of Akt reduced the extent of apoptosis induced by the antitumor drugs in Nalm-6 listed above. Activation of Akt did not induce expression of P-glycoprotein, a drug transporter that is capable of conferring multidrug resistance. Conclusion These results support the contention that Akt activation is a mechanism of chemotherapeutic resistance in B-pre ALL and suggest that Akt can be a therapeutic target for the treatment of relapsed or refractory pediatric B-pre ALL. Pediatr Blood Cancer 2012; 59: 83–89. © 2011 Wiley Periodicals, Inc.

Content maybe subject to copyright    Report

氏 名
授 与 し た 学 位
専 攻 分 野 の名 称
学 位 授 与 番 号
学 位 授 与 の 日付
学 位 授 与 の 要 件
学 位 論 文 題 目
森 下 直 人
博 士
医 学
博 甲第 4468 号
平成 24年 3月 2 3 日
医歯薬学総合研究科病態制御科学専攻
(学位 規則 第 4条 第 1項該 当)
ActivationofAktisAssociatedWithPoorPrognosis
and Chemotherapeutic Resistance in Pediatric
B‑PrecursorAcuteLymphoblasticLeukemia
(Akt
の活性化は､小児のB前駆細胞性急性 リンパ性 白血病に
おける予後不良と化学療法抵抗性に関連 している)
論 文 審 査 委 員
教授 谷 本 光音 教授 岩 月 啓 氏 教 授 杉 山 育
学 位 論 文 内 容 の 要 旨
pt3Kl
触 経路壮生存促進性の経路であり､リン酸化によるjkt浄財 ヒは腰痛細胞の成長におい
て重要な役割を果たしている｡しかし､小児
B前納 包性急性リンパ性白血病 (以下､RpreJqL)
の樹散生理におけるjgdの役割はいまだ不明の部分が多Llo今回､我々R 小児BpreAu.にお
けるj
kt活性化が臨床検査学的に分袖 壁学的にどのような意義を有するのか調べた｡
21名の小児 BpreAL患者を対象に行った飽和 勺検討では 骨髄倹体でjgd:が括性化している
患者では菅性化していない患者i3ヒべて化学馳 こ対する反応性が有意に乏しく､生命予後も有
意に悪いことが示 された
CIn trarBfectionによりAktを持続 的に活性化させたBpreA⊥細胞
株 恥 Im6細骨砂 を用いた基砲的検討では､抗脚 Jにより誘導されるアポ トーシスが有意に抑
制されることが示 された｡以上の結果より､
Bpre凡Lにおいて､jgdl措性化樹 臓 超抗性を
もたらす機構
の 1つであり､Jqdが†㈱ こ抵抗性を示 机 ＼児Bpre,qLL患都 こおいて治療標的
になりえることが示唆された｡
論 文 審 査 結 果 の 要 旨
本研究では､′J､児 B･preALLにおいて､骨髄検体で Aktが活性化 している患者 と活性
化が明らかでない患者 との予後を比較 した結果､
Aktの活性化が生存 とかかわっているこ
とを見出 した｡予後に影響を与える機構を解明するため
､B･preALL細胞株であるNalm‑6
株に活性化 した Akt遺伝子をせん孔法で導入 した細胞株に於いては､化学療法剤である
ステロイ ド､オンコビン､ア ドリアマイシン添加によるアポ トーシスが明 らかに抑制 され
ていることを見出 した｡この結果は､A
kt遺伝子の活性化経路がアポ トーシス抑制を介 し
て化学療法治療に対する抵抗性を獲得 していることを示 した新たな知見である｡
よって､本研究者は博士 (医学)の学位を得る資格があると認める｡
Citations
More filters
Journal Article•DOI•
Reversal effect of ST6GAL 1 on multidrug resistance in human leukemia by regulating the PI3K/Akt pathway and the expression of P-gp and MRP1

[...]

Hongye Ma1, Lei Cheng1, Keji Hao2, Yanping Li1, Xiaobo Song3, Huimin Zhou1, Li Jia1 •
Dalian Medical University1, Peking University2, University of Tromsø3
16 Jan 2014-PLOS ONE
TL;DR: It is postulated that ST6GAL1 is responsible for the development of MDR in human leukemia cells probably through medicating the activity of PI3K/Akt signaling and the expression of P-gp and MRP1.
Abstract: β-galactoside α2, 6-sialyltransferse gene (ST6GAL) family has two members, which encode corresponding enzymes ST6Gal I and ST6Gal II. The present atudy was to investigate whether and how ST6GAL family involved in multidrug resistance (MDR) in human leukemia cell lines and bone marrow mononuclear cells (BMMC) of leukemia patients. Real-time PCR showed a high expression level of ST6GAL1 gene in both MDR cells and BMMCs (*P<0.05). Alternation of ST6GAL1 levels had a significant impact on drug-resistant phenotype changing of K562 and K562/ADR cells both in vitro and in vivo. However, no significant changes were observed of ST6GAL2 gene. Further data revealed that manipulation of ST6GAL1 modulated the activity of phosphoinositide 3 kinase (PI3K)/Akt signaling and consequently regulated the expression of P-glycoprotein (P-gp, *P<0.05) and multidrug resistance related protein 1 (MRP1, *P<0.05), which are both known to be associated with MDR. Therefore we postulate that ST6GAL1 is responsible for the development of MDR in human leukemia cells probably through medicating the activity of PI3K/Akt signaling and the expression of P-gp and MRP1.

53 citations

Journal Article•DOI•
Targeting PI3K Signaling in Acute Lymphoblastic Leukemia

[...]

Vanessa Edna Sanchez1, Cydney Nichols2, Hye Na Kim1, Eun Ji Gang1, Yong-Mi Kim1 •
University of Southern California1, New York Medical College2
18 Jan 2019-International Journal of Molecular Sciences
TL;DR: The role of PI3K in normal hematopoietic cells, and in ALL is reviewed, with a focus on summarizing targeting strategies of PI2K in ALL.
Abstract: Adhesion of acute lymphoblastic leukemia (ALL) cells to bone marrow stroma cells triggers intracellular signals regulating cell-adhesion-mediated drug resistance (CAM-DR). Stromal cell protection of ALL cells has been shown to require active AKT. In chronic lymphocytic leukemia (CLL), adhesion-mediated activation of the PI3K/AKT pathway is reported. A novel FDA-approved PI3Kδ inhibitor, CAL-101/idelalisib, leads to downregulation of p-AKT and increased apoptosis of CLL cells. Recently, two additional PI3K inhibitors have received FDA approval. As the PI3K/AKT pathway is also implicated in adhesion-mediated survival of ALL cells, PI3K inhibitors have been evaluated preclinically in ALL. However, PI3K inhibition has yet to be approved for clinical use in ALL. Here, we review the role of PI3K in normal hematopoietic cells, and in ALL. We focus on summarizing targeting strategies of PI3K in ALL.

53 citations

Journal Article•DOI•
Modification of sialylation is associated with multidrug resistance in human acute myeloid leukemia

[...]

Hongye Ma1, Humin Zhou1, Xiaobo Song2, S Shi1, Jianing Zhang3, Li Jia1 •
Dalian Medical University1, University of Tromsø2, Dalian University of Technology3
05 Feb 2015-Oncogene
TL;DR: This study indicated that sialylation involved in the development of MDR of AML cells probably through ST3GAL5 or ST8SIA4 regulating the activity of PI3K/Akt signaling and the expression of P-gp and MRP1.
Abstract: Aberrant cell surface sialylation patterns have been shown to correlate with tumor progression and metastasis. However, the role of sialylation regulation of cancer multidrug resistance (MDR) remains poorly understood. This study investigated sialylation in modification on MDR in acute myeloid leukemia (AML). Using mass spectrometry (MS) analysis, the composition profiling of sialylated N-glycans differed in three pairs of AML cell lines. Real-time PCR showed the differential expressional profiles of 20 sialyltransferase (ST) genes in the both AML cell lines and bone marrow mononuclear cells (BMMCs) of AML patients. The expression levels of ST3GAL5 and ST8SIA4 were detected, which were overexpressed in HL60 and HL60/adriamycin-resistant (ADR) cells. The altered levels of ST3GAL5 and ST8SIA4 were found in close association with the MDR phenotype changing of HL60 and HL60/ADR cells both in vitro and in vivo. Further data demonstrated that manipulation of these two genes' expression modulated the activity of phosphoinositide-3 kinase (PI3K)/Akt signaling pathway and its downstream target thus regulated the proportionally mutative expression of P-glycoprotein (P-gp) and MDR-related protein 1 (MRP1), both of which are known to be involved in MDR. Blocking the PI3K/Akt pathway by its specific inhibitor LY294002 or by Akt small interfering RNA resulted in the reduced chemosensitivity of HL60/ADR cells. Therefore, this study indicated that sialylation involved in the development of MDR of AML cells probably through ST3GAL5 or ST8SIA4 regulating the activity of PI3K/Akt signaling and the expression of P-gp and MRP1.

51 citations

Journal Article•DOI•
Phase I study of UCN-01 and perifosine in patients with relapsed and refractory acute leukemias and high-risk myelodysplastic syndrome

[...]

Ivana Gojo1, Ivana Gojo2, Alexander E. Perl3, Selina M. Luger3, Maria R. Baer1, Kelly J. Norsworthy1, Kenneth S. Bauer4, Kenneth S. Bauer1, Michael L. Tidwell1, Stephanie Fleckinger1, Martin Carroll3, Edward A. Sausville1 •
University of Maryland Marlene and Stewart Greenebaum Cancer Center1, Johns Hopkins University2, University of Pennsylvania3, University of Maryland, Baltimore4
27 Feb 2013-Investigational New Drugs
TL;DR: UCN-01 and perifosine can be safely administered, but this regimen lacked clinical efficacy and the approach may have failed because of insufficient Akt inhibition in vivo.
Abstract: Background The PI3K-Akt pathway is frequently activated in acute leukemias and represents an important therapeutic target. UCN-01 and perifosine are known to inhibit Akt activation. Methods The primary objective of this phase I study was to determine the maximum tolerated dose (MTD) of UCN-01 given in combination with perifosine in patients with advanced acute leukemias and myelodysplastic syndrome. Secondary objectives included safety, pharmacokinetics, pharmacodynamics, and efficacy. Perifosine 150 mg every 6 h was given orally on day 1 followed by 100 mg once a day continuously in 28-day cycles. UCN-01 was given intravenously over 3 h on day 4 at three dose levels (DL1 = 40 mg/m2; DL2 = 65 mg/m2; DL3 = 90 mg/m2). Results Thirteen patients were treated (DL1, n = 6; DL2, n = 4; DL3, n = 3) according to a traditional “3 + 3” design. Two patients at the DL3 experienced dose-limiting toxicity including grade 3–4 pericardial effusion, hypotension, hyperglycemia, hyperkalemia, constitutional symptoms and grade 5 pneumonitis. Other frequent toxicities were grade 1–2 nausea, diarrhea, vomiting, fatigue and hyperglycemia. The MTD was determined to be UCN-01 65 mg/m2 with perifosine 100 mg a day. No appreciable direct Akt inhibition could be demonstrated in patients’ mononuclear cells using Western blot, however, reduced phosphorylation of the downstream target ribosomal protein S6 in leukemic blasts was noted by intracellular flow cytometry. No objective responses were observed on this study. Conclusion UCN-01 and perifosine can be safely administered, but this regimen lacked clinical efficacy. This approach may have failed because of insufficient Akt inhibition in vivo.

49 citations

Journal Article•DOI•
Control of B lymphocyte development and functions by the mTOR signaling pathways.

[...]

Terri Iwata1, Julita A. Ramírez-Komo1, Heon Park1, Brian M. Iritani1•
University of Washington1
01 Jun 2017-Cytokine & Growth Factor Reviews
TL;DR: The basic mechanisms of mTOR signaling are summarized; what is known about the roles of m TORC1, mTORC2, and the Folliculin/Fnip1 pathway in B cell development and functions are described; and current clinical approaches for targeting mTOR in B Cell neoplasms are outlined.

41 citations

References
More filters
Journal Article•DOI•
AKT/PKB signaling: navigating downstream.

[...]

Brendan D. Manning1, Lewis C. Cantley2•
Harvard University1, Beth Israel Deaconess Medical Center2
29 Jun 2007-Cell
TL;DR: Those Akt substrates that are most likely to contribute to the diverse cellular roles of Akt, which include cell survival, growth, proliferation, angiogenesis, metabolism, and migration are discussed.

5,505 citations

"Activation of Akt is associated wit..." refers background in this paper

  • ...Akt binds to phosphatidylinositol-3,4,5-triphosphate on the plasma membrane where Akt is activated by phosphorylation at threonine308 (T308) and serine473 (S473) residues....

    [...]

  • ...Antibodies against Akt and PAkt on S473 and HRP-linked secondary antibody were purchased from Cell Signaling Technology Inc. (Irvine, CA) and antibodies against b-actin and Myc tag from Santa Cruz Biotechnology Inc. and Millipore Corp. (Billerica, MA), respectively....

    [...]

  • ...30-Phosphoinositide-dependent kinase 1 phosphorylates the activation loop of Akt at T308, although the mammalian target of rapamycin complex 2 phosphorylates Akt at the hydrophobic motif S473 [7]....

    [...]

Journal Article•DOI•
The activation of Akt/PKB signaling pathway and cell survival

[...]

Gang Song1, Gaoliang Ouyang1, Shideng Bao2, Shideng Bao1•
Xiamen University1, Duke University2
01 Jan 2005-Journal of Cellular and Molecular Medicine
TL;DR: Akt/PKB plays important roles in the signaling pathways in response to growth factors and other extracellular stimuli to regulate several cellular functions including nutrient metabolism, cell growth, apoptosis and survival.
Abstract: Akt/PKB is a serine/threonine protein kinase that functions as a critical regulator of cell survival and proliferation. Akt/PKB family comprises three highly homologous members known as PKBalpha/Akt1, PKBbeta/Akt2 and PKBgamma/Akt3 in mammalian cells. Similar to many other protein kinases, Akt/PKB contains a conserved domain structure including a specific PH domain, a central kinase domain and a carboxyl-terminal regulatory domain that mediates the interaction between signaling molecules. Akt/PKB plays important roles in the signaling pathways in response to growth factors and other extracellular stimuli to regulate several cellular functions including nutrient metabolism, cell growth, apoptosis and survival. This review surveys recent developments in understanding the molecular mechanisms of Akt/PKB activation and its roles in cell survival in normal and cancer cells.

1,811 citations

"Activation of Akt is associated wit..." refers background in this paper

  • ...We assume that other pathways aside from P-glycoprotein play an important role in MDR in Nalm-6/Akt, such as expression of another ATP-binding cassette transporter, inhibition of BAD, inhibition of Caspase-9, activation of NF-kB, inhibition of p53, and so forth [6,24]....

    [...]

  • ...Activation of the PI3K/Akt pathway induces activation of nuclear factor-kB and Bcl-2 and inhibition of Bcl-2 antagonist of cell death (BAD), Caspase-9 and p53 [6]....

    [...]

  • ...The phosphoinositide 3-kinase (PI3K)/Akt signaling pathway is a prototypic survival pathway that plays a central role in diverse cellular functions including insulin metabolism, protein synthesis, proliferation, and apoptosis [5,6]....

    [...]

Journal Article•DOI•
P-glycoprotein: from genomics to mechanism

[...]

Suresh V. Ambudkar, Chava Kimchi-Sarfaty, Zuben E. Sauna, Michael M. Gottesman
20 Oct 2003-Oncogene
TL;DR: A model for how ATP energizes transfer of substrates from these binding sites on P-gp to the outside of the cell is proposed, which accounts for the apparent stoichiometry of two ATPs hydrolysed per molecule of drug transported.
Abstract: Resistance to chemically different natural product anti-cancer drugs (multidrug resistance, or MDR) results from decreased drug accumulation, resulting from expression of one or more ATP-dependent efflux pumps. The first of these to be identified was P-glycoprotein (P-gp), the product of the human MDR1 gene, localized to chromosome 7q21. P-gp is a member of the large ATP-binding cassette (ABC) family of proteins. Although its crystallographic 3-D structure is yet to be determined, sequence analysis and comparison to other ABC family members suggest a structure consisting of two transmembrane (TM) domains, each with six TM segments, and two nucleotide-binding domains. In the epithelial cells of the gastrointestinal tract, liver, and kidney, and capillaries of the brain, testes, and ovaries, P-gp acts as a barrier to the uptake of xenobiotics, and promotes their excretion in the bile and urine. Polymorphisms in the MDR1 gene may affect the pharmacokinetics of many commonly used drugs, including anticancer drugs. Substrate recognition of many different drugs occurs within the TM domains in multiple-overlapping binding sites. We have proposed a model for how ATP energizes transfer of substrates from these binding sites on P-gp to the outside of the cell, which accounts for the apparent stoichiometry of two ATPs hydrolysed per molecule of drug transported. Understanding of the biology, genetics, and biochemistry of P-gp promises to improve the treatment of cancer and explain the pharmacokinetics of many commonly used drugs.

985 citations

"Activation of Akt is associated wit..." refers background in this paper

  • ...P-glycoprotein is a member of ATP-binding cassette transporters that extract antitumor drugs from cytoplasm and which reduce intracellular drug concentrations to sub-lethal levels [20,21]....

    [...]

  • ...The expression of P-glycoprotein, encoded by the MDR1 gene, is often associated with MDR in tumor cells [20,27]....

    [...]

  • ...In patients with leukemias, lymphomas, and multiple myeloma, P-glycoprotein expression, which is very low at presentation, is occasionally enhanced during administration of long-term chemotherapy or when the disease recurs [20]....

    [...]

Journal Article•DOI•
Survival signalling by Akt and eIF4E in oncogenesis and cancer therapy.

[...]

Hans-Guido Wendel1, Elisa de Stanchina1, Jordan S. Fridman2, Jordan S. Fridman1, Abba Malina3, Sagarika Ray1, Scott C. Kogan4, Carlos Cordon-Cardo5, Jerry Pelletier3, Scott W. Lowe1 •
Cold Spring Harbor Laboratory1, Incyte2, McGill University3, University of California, San Francisco4, Memorial Sloan Kettering Cancer Center5
18 Mar 2004-Nature
TL;DR: It is shown that Akt promotes tumorigenesis and drug resistance by disrupting apoptosis, and that disruption of Akt signalling using the mTOR inhibitor rapamycin reverses chemoresistance in lymphomas expressing Akt, but not in those with other apoptotic defects.
Abstract: Evading apoptosis is considered to be a hallmark of cancer, because mutations in apoptotic regulators invariably accompany tumorigenesis. Many chemotherapeutic agents induce apoptosis, and so disruption of apoptosis during tumour evolution can promote drug resistance. For example, Akt is an apoptotic regulator that is activated in many cancers and may promote drug resistance in vitro. Nevertheless, how Akt disables apoptosis and its contribution to clinical drug resistance are unclear. Using a murine lymphoma model, we show that Akt promotes tumorigenesis and drug resistance by disrupting apoptosis, and that disruption of Akt signalling using the mTOR inhibitor rapamycin reverses chemoresistance in lymphomas expressing Akt, but not in those with other apoptotic defects. eIF4E, a translational regulator that acts downstream of Akt and mTOR, recapitulates Akt's action in tumorigenesis and drug resistance, but is unable to confer sensitivity to rapamycin and chemotherapy. These results establish Akt signalling through mTOR and eIF4E as an important mechanism of oncogenesis and drug resistance in vivo, and reveal how targeting apoptotic programmes can restore drug sensitivity in a genotype-dependent manner.

949 citations

"Activation of Akt is associated wit..." refers background in this paper

  • ...Furthermore, murine models of Akt activation have shown that constitutively active Akt causes myeloproliferative disease [30], promotes drug resistance, and shortens tumor-free survival [25]....

    [...]

Journal Article•DOI•
The Akt-mTOR tango and its relevance to cancer.

[...]

Nissim Hay1•
University of Illinois at Chicago1
01 Sep 2005-Cancer Cell
TL;DR: Two recent studies used mouse genetics to assess the roles of PTEN and TSC2 in cancer, underscoring the importance of Akt-mTOR interplay for cancer progression and therapy.

767 citations

"Activation of Akt is associated wit..." refers background in this paper

  • ...The phosphoinositide 3-kinase (PI3K)/Akt signaling pathway is a prototypic survival pathway that plays a central role in diverse cellular functions including insulin metabolism, protein synthesis, proliferation, and apoptosis [5,6]....

    [...]

  • ...The PI3K/Akt signaling pathway controls the expression and function of many proteins that are necessary for cell survival, growth, and anti-apoptosis [5]....

    [...]

Related Papers (5)
Multidrug resistance-associated protein 1 expression is under the control of the phosphoinositide 3 kinase/Akt signal transduction network in human acute myelogenous leukemia blasts

[...]

11 Jan 2007-Leukemia
P. L. Tazzari, Alessandra Cappellini, Francesca Ricci, Camilla Evangelisti, Veronica Papa, Tiziana Grafone, Giovanni Martinelli, Roberto Conte, Lucio Cocco, James A. McCubrey, Alberto M. Martelli, Alberto M. Martelli 
Direct reversal of glucocorticoid resistance by AKT inhibition in acute lymphoblastic leukemia

[...]

09 Dec 2013-Cancer Cell
Erich Piovan, Erich Piovan, Jiyang Yu, Valeria Tosello, Daniel Herranz, Alberto Ambesi-Impiombato, Ana Carolina Da Silva, Marta Sanchez-Martin, Arianne Perez-Garcia, Isaura Rigo, Mireia Castillo, Stefano Indraccolo, Justin R. Cross, Elisa de Stanchina, Elisabeth Paietta, Elisabeth Paietta, Janis Racevskis, Janis Racevskis, Jacob M. Rowe, Martin S. Tallman, Giuseppe Basso, Jules P.P. Meijerink, Carlos Cordon-Cardo, Andrea Califano, Adolfo A. Ferrando 
Rapamycin is active against B-precursor leukemia in vitro and in vivo, an effect that is modulated by IL-7-mediated signaling.

[...]

09 Dec 2003-Proceedings of the National Academy of Sciences of the United States of America
Valerie I. Brown, Junjie Fang, Keith Alcorn, Rosalind Barr, Jenny M. Kim, Robert Wasserman, Stephan A. Grupp 
PTEN posttranslational inactivation and hyperactivation of the PI3K/Akt pathway sustain primary T cell leukemia viability.

[...]

03 Nov 2008-Journal of Clinical Investigation
Ana Elisa Bauer de Camargo Silva, J. Andrés Yunes, Bruno A. Cardoso, Leila R. Martins, Patrícia Y. Jotta, Miguel Abecasis, Alexandre E. Nowill, Nick R. Leslie, Angelo A. Cardoso, João T. Barata 
Gene expression-based chemical genomics identifies rapamycin as a modulator of MCL1 and glucocorticoid resistance

[...]

01 Oct 2006-Cancer Cell
Guo Wei, Guo Wei, David Twomey, David Twomey, Justin Lamb, Krysta D. Schlis, Jyoti Agarwal, Ronald W. Stam, Joseph T. Opferman, Stephen E. Sallan, Monique L. den Boer, Rob Pieters, Todd R. Golub, Scott A. Armstrong