Design, synthesis, and evaluation of small molecule Hsp90 probes

Taldone, Tony; Zatorska, Danuta; Patel, Pallav D.; Zong, Hong; Rodina, Anna; Ahn, James H.; Moulick, Kamalika; Guzmán, Mónica L.; Chiosis, Gabriela

doi:10.1016/j.bmc.2011.03.013

Cited by 47 publications

(49 citation statements)

References 41 publications

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“…17-DMAG was purchased from Sigma. HSP90 bait (PU-H71 beads) 21 , HSP70 bait (YK beads) 22 , biotinylated YK (YK-biotin) 22 , fluorescently labelled PU-H71 (PU-FITC) 23 , the control derivatives PU-TEG and PU-FITC9 (ref. 24), and the radiolabelled PU-H71-derivative 124 I-PU-H71 (ref.…”

Section: Methodsmentioning

confidence: 99%

The epichaperome is an integrated chaperome network that facilitates tumour survival

Rodina

Wang

Yan

et al. 2016

Nature

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Transient, multi-protein complexes are important facilitators of cellular functions. This includes the chaperome, an abundant protein family comprising chaperones, co-chaperones, adaptors, and folding enzymes—dynamic complexes of which regulate cellular homeostasis together with the protein degradation machinery1–6. Numerous studies have addressed the role of chaperome members in isolation, yet little is known about their relationships regarding how they interact and function together in malignancy7–17. As function is probably highly dependent on endogenous conditions found in native tumours, chaperomes have resisted investigation, mainly due to the limitations of methods needed to disrupt or engineer the cellular environment to facilitate analysis. Such limitations have led to a bottleneck in our understanding of chaperome-related disease biology and in the development of chaperome-targeted cancer treatment. Here we examined the chaperome complexes in a large set of tumour specimens. The methods used maintained the endogenous native state of tumours and we exploited this to investigate the molecular characteristics and composition of the chaperome in cancer, the molecular factors that drive chaperome networks to crosstalk in tumours, the distinguishing factors of the chaperome in tumours sensitive to pharmacologic inhibition, and the characteristics of tumours that may benefit from chaperome therapy. We find that under conditions of stress, such as malignant transformation fuelled by MYC, the chaperome becomes biochemically ‘rewired’ to form a network of stable, survival-facilitating, high-molecular-weight complexes. The chaperones heat shock protein 90 (HSP90) and heat shock cognate protein 70 (HSC70) are nucleating sites for these physically and functionally integrated complexes. The results indicate that these tightly integrated chaperome units, here termed the epichaperome, can function as a network to enhance cellular survival, irrespective of tissue of origin or genetic background. The epichaperome, present in over half of all cancers tested, has implications for diagnostics and also provides potential vulnerability as a target for drug intervention.

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Section: Methodsmentioning

confidence: 99%

The epichaperome is an integrated chaperome network that facilitates tumour survival

Rodina

Wang

Yan

et al. 2016

Nature

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230

531

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“…Cells were cultured in RPMI-1640 media (Invitrogen) supplemented with 10% heat inactivated fetal bovine serum and gentamicin 50ug/mL (Sigma-Aldrich). PU-H71, PU-H71-beads, and control beads were synthesized and prepared at Memorial Sloan Kettering Cancer Center as previously reported(13,14). Idelalisib (CAL-101), IPI-145, BKM120 (15), BEZ-235 (16), ganetespib (17), and BIIB-021 (18) were purchased from Selleck Chemicals.…”

Section: Methodsmentioning

confidence: 99%

Inhibition of Hsp90 Suppresses PI3K/AKT/mTOR Signaling and Has Antitumor Activity in Burkitt Lymphoma

Giulino‐Roth

Besien

Dalton

et al. 2017

Molecular Cancer Therapeutics

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Heat shock protein 90 (Hsp90) is a molecular chaperone that protects proteins, including oncogenic signaling complexes, from proteolytic degradation. PU-H71 is a next-generation Hsp90 inhibitor that preferentially targets the functionally distinct pool of Hsp90 present in tumor cells. Tumors that are driven by the MYC oncoprotein may be particularly sensitive to PU-H71 due to the essential role of Hsp90 in the epichaperome which maintains the malignant phenotype in the setting of MYC. Burkitt lymphoma (BL) is an aggressive B-cell lymphoma characterized by MYC dysregulation. In the current study we evaluated Hsp90 as a potential therapeutic target in Burkitt lymphoma. We found that primary BL tumors overexpress Hsp90 and that Hsp90 inhibition has anti-tumor activity in-vitro and in-vivo, including potent activity in a patient derived xenograft model of BL. To evaluate the targets of PU-H71 in BL we performed high affinity capture followed by proteomic analysis using mass spectrometry. We found that Hsp90 inhibition targets multiple components of PI3K/AKT/mTOR signaling, highlighting the importance of this pathway in BL. Lastly, we found that the anti-lymphoma activity of PU-H71 is synergistic with dual PI3K/mTOR inhibition in-vitro and in-vivo. Overall this work provides support for Hsp90 as a therapeutic target in BL and suggests the potential for combination therapy with PU-H71 and inhibitors of PI3K/mTOR.

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“…Client proteins are inferred by co-isolation. 24, 25 However, in vivo, many clients undergoing folding are thought to be weakly associated with Hsp90, potentially complicating their identification using these approaches. Therefore, it seemed one could benefit from the use of a highly selective affinity probe that would allow for efficient Hsp90 release under mild conditions while discriminating against non-specifically associated proteins that typically bind to resin surfaces.…”

Section: Introductionmentioning

confidence: 99%

A highly selective Hsp90 affinity chromatography resin with a cleavable linker

Hughes

Barrott

Carlson

et al. 2012

Bioorganic & Medicinal Chemistry

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Over 200 proteins have been identified that interact with the protein chaperone Hsp90, a recognized therapeutic target thought to participate in non-oncogene addiction in a variety of human cancers. However, defining Hsp90 clients is challenging because interactions between Hsp90 and its physiologically relevant targets involve low affinity binding and are thought to be transient. Using a chemo-proteomic strategy, we have developed a novel orthogonally cleavable Hsp90 affinity resin that allows purification of the native protein and is quite selective for Hsp90 over its immediate family members, GRP94 and TRAP 1. We show that the resin can be used under low stringency conditions for the rapid, unambiguous capture of native Hsp90 in complex with a native client. We also show that the choice of linker used to tether the ligand to the insoluble support can have a dramatic effect on the selectivity of the affinity media.

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Design, synthesis, and evaluation of small molecule Hsp90 probes

Cited by 47 publications

References 41 publications

The epichaperome is an integrated chaperome network that facilitates tumour survival

The epichaperome is an integrated chaperome network that facilitates tumour survival

Inhibition of Hsp90 Suppresses PI3K/AKT/mTOR Signaling and Has Antitumor Activity in Burkitt Lymphoma

A highly selective Hsp90 affinity chromatography resin with a cleavable linker

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