MALE CONTRACEPTION UPDATE

July 2008
Volume 3, Issue 7

In this issue

Summaries of new peer-reviewed publications

RISUG update: An interview with Dr. Sharma of the ICMR

Press coverage

Calendar of events

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The Male Contraception Update e-newsletter is brought to you every month by the Male Contraception Information Project and the Male Contraception Coalition.

Summaries of new peer-reviewed publications

Reviews
"To analyze the impact of commonly used drugs on male fertility, we assessed the clinical characteristics of patients with impaired semen quality while they were taking medication for chronic diseases and after switching therapies... Our results confirm the potential fertility hazards of commonly used drugs and their reversibility. Moreover, after switching medication, drug-induced asthenozoospermia was cured more rapidly than oligozoospermia, suggesting that further delineation of such differences may help to elucidate mechanisms of spermatogenesis and might facilitate the development of non-hormonal male contraceptive agents."
The impact of commonly prescribed drugs on male fertility.
Hayashi T, Miyata A, Yamada T.
Hum Fertil (Camb). 2008 Jun 17:1-6. [Epub ahead of print]
PMID: 18608524

Cell adhesion targets
"The Par3/Par6/aPKC and the CRB3/Pals1/PATJ polarity complexes are involved in regulating apical ectoplasmic specialization (ES) and blood-testis barrier (BTB) restructuring in the testis... When rats were treated with adjudin to induce apical ES restructuring without compromising the BTB, Par6 staining virtually disappeared at the apical ES in misaligned spermatids before their depletion... When either Par6 or Par3 was knocked down by RNAi in Sertoli cell epithelium, a significant loss of the corresponding protein by approximately 60% in cells vs. controls was detected, alongside with a decline in aPKC after Par6, but not Par3, knockdown. This Par3 or Par6 knockdown also led to a transient loss of selected BTB proteins at the cell-cell interface, thereby compromising the BTB integrity."
Par3/Par6 polarity complex coordinates apical ectoplasmic specialization and blood-testis barrier restructuring during spermatogenesis.
Wong EW, Mruk DD, Lee WM, Cheng CY.
Proc Natl Acad Sci U S A. 2008 Jul 15;105(28):9657-62. Epub 2008 Jul 10.
PMID: 18621709

"The mechanism(s) that regulate and coordinate the events of spermiation and blood-testis barrier (BTB) restructuring in the seminiferous epithelium that occur concurrently at stage VIII of the seminiferous epithelial cycle of spermatogenesis are unknown. In this report, fragments derived from the laminin complex composed of laminin alpha3, beta3, and gamma3 chains (laminin-333) at the apical ectoplasmic specialization (apical ES) were shown to modulate BTB dynamics directly and/or indirectly via hemidesmosome."
An autocrine axis in the testis that coordinates spermiation and blood-testis barrier restructuring during spermatogenesis.
Yan HH, Mruk DD, Wong EW, Lee WM, Cheng CY.
Proc Natl Acad Sci U S A. 2008 Jul 1;105(26):8950-5. Epub 2008 Jun 25.
PMID: 18579774

Immunological approaches
"Is it possible to deliver therapeutic agents to testis through specific targeting? Immunoliposomes are designed by incorporating antibodies to lactate dehydrogenase-C4 (LDH-C(4)), which is the product of a testis specific gene... It is observed that LDHC(4)-immunoliposomes are able to discriminate and recognize antigens on spermatozoa and testes both in vitro and in vivo. Specific targeting through LDH-C(4) appears to be a feasible strategy for delivering therapeutic as well as anti-spermatogenic agents to testis."
Testis specific lactate dehydrogenase as target for immunoliposomes.
Dutta RC, Goldberg E.
Am J Reprod Immunol. 2008 Jul;60(1):26-32.
PMID: 18593435

Endocrinological approaches
"It is disappointing that both pharmaceutical companies involved in this study, and the most active in the MHC development, have since decided to withdraw from the field. Their reasons are sure to be multifactorial but probably include the perceived profitability of male hormonal contraception in the context of a shrinking global market and an increasingly adversarial medicolegal climate. To this end, the public needs to clearly articulate its wishes because there is a clear disconnect between what academic clinicians hear from disgruntled couples and what the pharmaceutical industry and governments believe!"
Male hormonal contraception: so near and yet so far.
Liu PY, McLachlan RI.
J Clin Endocrinol Metab. 2008 Jul;93(7):2474-6.
PMID: 18617702

Proteomic / genomic supporting research
"This study investigates the role of caspase 2 in apoptotic signaling of nonhuman primate male germ cells triggered by mild testicular hyperthermia, testosterone (T) implants, or by combined interventions... Most notably, active caspase 2 immunoreactivity was detected only in those germ cells susceptible to apoptosis... Caspase 2 inhibition significantly (P < 0.05) prevented such heat-induced germ cell apoptosis. The protection offered by the caspase 2 inhibitor occurred upstream of mitochondria, involving suppression of mitogen-activated protein kinase (MAPK) 14 activation and inducible nitric oxide synthase (NOS2) induction and, in turn, suppression of cytochrome c-mediated death pathway."
Role of Caspase 2 in Apoptotic Signaling in Primate and Murine Germ Cells.
Johnson C, Jia Y, Wang C, Lue YH, Swerdloff RS, Zhang XS, Hu ZY, Li YC, Liu YX, Sinha-Hikim AP.
Biol Reprod. 2008 Jul 9. [Epub ahead of print]
PMID: 18614702

"The enzyme Dicer1 is required for miRNA processing and mouse knockouts of Dicer1 are embryonic lethal before E7.5. To examine the function of miRNAs specifically in the germline we used a mouse model which expresses Cre recombinase from the TNAP locus and a floxed Dicer1 conditional allele. Removal of Dicer1 from germ cells resulted in male infertility. Germ cells were present in adult testes, but few tubules contained elongating spermatids. Germ cells that did differentiate to elongating spermatids exhibited abnormal morphology and motility. Rarely, sperm lacking Dicer1 could fertilize wild type eggs to generate viable offspring. These results show that Dicer1 and miRNAs are essential for proper differentiation of the male germline."
Dicer1 Is Required for Differentiation of the Mouse Male Germline.
Maatouk DM, Loveland KL, McManus MT, Moore K, Harfe BD.
Biol Reprod. 2008 Jul 16. [Epub ahead of print]
PMID: 18633141

"In this study, we used 2D-PAGE and MALDI-TOF/TOF technology to construct a comparative proteome profile for mouse testis at specific time points (days 0, 7, 14, 21, 28, and 60 postpartum). We identified 362 differential protein spots corresponding to 257 different proteins. Further cluster analysis revealed 6 expression patterns, and bioinformatics analysis revealed that each pattern was related to many specific cell processes. Among them, 28 novel proteins with unknown functions neither in somatic cells nor germ cells were identified, 8 of which were found to be uniquely or highly expressed in mouse testes via comparison with the GNF SymAtlas database."
Construction of a Proteome Profile and Functional Analysis of the Proteins Involved in the Initiation of Mouse Spermatogenesis.
Huang XY, Guo XJ, Shen J, Wang YF, Chen L, Xie J, Wang NL, Wang FQ, Zhao C, Huo R, Lin M, Wang X, Zhou ZM, Sha JH.
J Proteome Res. 2008 Jun 27. [Epub ahead of print]
PMID: 18582094

A novel testis-specific gene termed mtIQ1 (GenBank Accession No. DQ153246) was identified by digital differential display. Sequence analysis revealed that mtIQ1 protein is a new member of calmodulin (CaM) binding protein families with conserved Ile and Gln residues (IQ motif). RT-PCR and Northern blot analysis revealed that a 0.9 kb mtIQ1 transcript was only expressed in adult mouse testis and not expressed in nine other tissues... Results of in situ hybridization assay confirmed that mtIQ1 was expressed in seminiferous tubules, more precisely in spermatocytes."
Molecular cloning and expression profile analysis of a novel mouse testis-specific expression gene mtIQ1.
Nie D, Yang X, Yankai Z.
Mol Biol Rep. 2008 Jun 27. [Epub ahead of print]
PMID: 18584305

"The purpose of this study was to examine the availability, target cell types and efficiency of RNAi in the mouse seminiferous epithelium...Sertoli cells were the main transfected cells. A reduction of about 40% in the level of EGFP [(enhanced green fluorescent] protein was detected in cells successfully transfected both in vivo and in vitro. However, the efficiency of in vivo transfection was low. In adult seminiferous epithelial cells, in vivo post-transcriptional gene silencing mediated by RNAi via shRNA is efficient in Sertoli cells. Similar levels of RNAi were detected both in vivo and in vitro. This also indicates that Sertoli cells have the necessary silencing machinery to repress the expression of endogenous genes via RNAi."
Gene silencing by RNAi in mouse Sertoli cells.
Gonzalez-Gonzalez E, Lopez-Casas PP, Del Mazo J.
Reprod Biol Endocrinol. 2008 Jul 11;6(1):29. [Epub ahead of print]
PMID: 18620581

Gamete binding / fusion targets
"To investigate the molecular regulation of spermatogenesis in vivo, we used differential display RT-PCR to identify testis-specific genes in a retinol-supplemented vitamin A deficiency (VAD) rat model and identified the VAD1.2 (acrosome-expressing protein 2, AEP2) gene, which was expressed strongly in the rat testis from postnatal day 32 to adult stage... VAD1.2 transcript was abundantly expressed in the rat seminiferous tubules at stage VIII-XII and the protein was detected in the acrosome region of the round and elongated spermatids of mouse, human, monkey and pig. VAD1.2 co-localized with lectin-PNA to the acrosome region of spermatids... VAD1.2 may be involved in acrosome formation during spermiogenesis."
Characterization of an acrosome protein VAD1.2/AEP2 which is differentially expressed in spermatogenesis.
Lee KF, Tam YT, Zuo Y, Cheong AW, Pang RT, Lee NP, Shum CK, Tam PC, Cheung AN, Yang ZM, S B Yeung W, Luk JM.
Mol Hum Reprod. 2008 Jul 10. [Epub ahead of print]
PMID: 18621766

Semen parameter standardization
"Comparison of methods that estimate viability of human spermatozoa revealed that wet preparations (whether using positive or negative phase contrast microscopy) generated significantly higher percentages of non-viable cells than air-dried, eosin-nigrosin smears. Only with the latter method did the sum of motile (live) and stained (dead) preparations never exceed 100%, making this the method of choice for sperm viability estimates."
Method-Related Estimates of Sperm Viability.
Cooper TG, Hellenkemper B.
J Androl. 2008 Jul 3. [Epub ahead of print]
PMID: 18599882

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RISUG update: An interview with Dr. Sharma of the ICMR

Dr. Sharma

This month we continue our series on male contraceptive research outside the U.S. Dr. R. S. Sharma is Deputy Director General (Senior Grade) & Scientist, Division of Reproductive Health and Nutrition at the Indian Council of Medical Research. He's a busy man, with oversight over, among other things, both male contraceptive research and national IVF standards. We caught up with him this month for an update on RISUG, the long-acting, non-hormonal intravasal injectable male contraceptive currently in Phase III clinical trial stage.

Male Contraception Information Project: How is the RISUG clinical trial proceeding?

Dr. R.S. Sharma: We are currently running two studies: one a follow-up study (“Comparative long-term retrospective follow up study of RISUG injected and vasectomized (NSV) subjects”) and the second a Phase III clinical trial (“Phase III Clinical Trial with an Intravasal Injectable Male Contraceptive – RISUG®”).

In the follow-up study, we are following all the subjects enrolled during the limited Phase III trial, in which 139 subjects were injected during 2001-2002. We are working on tracking down the subjects. So far we have identified about 66 subjects. Based on the criteria defined in the protocol, around 44 Non Scalpel Vasectomy (NSV) subjects have also been identified and followed for clinical and semen examination. No serious adverse clinical symptoms have been noticed in any of the subjects in either group 5 to 7 years after RISUG injection. There have been no intact sperm in any of the follow-up subjects even after 5 to 7 years.

In the new Phase III trial, we have started the study at four centers around the country, and have injected RISUG in about 64 subjects; the goal will be to enroll a total of 500 subjects. It is a slow process to get these studies started, and we had to complete many steps: we've done a workshop to train the paramedical staff, a workshop to train the investigators in injection procedures, and more.

I recently got the 6-month report on this study, at which time 64 injection procedures had been performed. The data received is under analysis, but it appears that there were no intact sperm in any of the 64 subjects.

MCIP: No intact sperm in any of the 64 subjects in the new study?

Sharma: Yes, that is correct. Although the detailed analysis is still going on, we are also quite encouraged that we have not come across any major side effects. This time, of the type of the swelling and pain noted occasionally in the Limited Phase III trial, the pain was not there, and the swelling was much less. I think the professionally manufactured drug may be helping - the drug was manufactured by a pharmaceutical company rather than in the laboratory, and that may be making a difference. But also, it may be due to training: we have designated a master trainer (Dr. H.C. Das, who participated in earlier trials) who is training all the new investigators in the injection procedure under the supervision of experts and also supervising the investigators in the beginning stages until they have perfected it.

MCIP: Those are incredible results! When might we see publication of this data? People are very excited about the possibility of a long-acting male contraceptive, but so far all the claims of long-term efficacy and reversibility have been based on animal data plus informal news from the men who stayed in touch. Why not publish data on the men from the one study site while you track down the others?

Sharma: Sixty six subjects is good, and this is certainly encouraging information, but we will feel more confident about having significant data if we can report on at least 100 of the 139 subjects. I have thought about publishing the interim results of the one study, but we have been stretched thin. There is another factor: we are following the NSV [No-Scalpel Vasectomy] group from the same time and the same duration of use, and we are focused on publishing the full comparison of the data sets. The results of the comparison between the two will be very important.

MCIP: What about reversibility? This is another issue where we've had to extrapolate from monkey studies. Reversibility is the key to opening up RISUG use to a whole generation of men not ready for permanent vasectomy. But there's always that nagging issue in the back of our minds: sure, monkeys can use it for a year and a half and still regain fertility upon reversal, and sure, publications show men's prostate markers and reproductive organs are normal even after years of use; but does that mean a man could really father a child after 5 or 10 years of use?

Sharma: Yes, reversibility is the issue. It's very, very important for us to look at this.

We are planning to develop a protocol to conduct reversal in a few subjects from the Phase I trial [17 or more years of use], Phase II trial [about 12 years of use], and/or Limited Phase III trial [6-7 years of use]. The monitoring committee has directed us to start this activity. Even after finalizing a joint protocol on reversibility, it will still be quite challenging: It's not in our hands to know how many men will accept reversal. We'll try our best, though, to get enough. This is certainly a very important issue we need to address.

MCIP: What are the next steps in this research?

Sharma: One of the most important steps is conducting a reversibility study, and extending the ongoing Phase III trial to 6 more centers.

MCIP: What do you see as a major hurdle or concern for RISUG?

Sharma: The enrollment of the subjects in the Phase III clinical trial is very poor. In India, male participation is poor even for vasectomy or NSV compared to female sterilization. So our main issue is to attract the male to accept this contraception option. It takes time to convince people, but we're taking steps such as placing ads in the newspaper and holding information meetings.

MCIP: But we were under the impression that there is a waitlist for injections, and the obstacle is steady supply of the RISUG material?

Sharma: That was an issue before, but it has been solved.

MCIP: Surely the well-educated urban men would be among the first adopters of this new technology, and publicity in the press would result in a great demand? Is there any role for publicity?

Sharma: Perhaps there could be a role for a joint effort in this area. I have been contacted by quite a few men from outside India who want to get RISUG, but the government hasn't permitted foreigners to participate in the trial, so we have to turn them down.

MCIP: Is there any other male contraceptive research you are excited about or would like to comment on?

Sharma: I am currently concentrating on RISUG, but I am also interested in promoting data-driven condom development. At the government's request, I was the first person to collect specifications [for appropriate size and shape] for condoms, and I found state-to-state variation within the country of India. If we want condoms to be effective and accepted, then we need to develop condoms to the correct specifications.

If my plate weren't so full, I would like to be collecting data for the regions that we didn't study, and testing the efficacy and acceptability of new condoms developed based on these new specifications. Condoms are extremely important, with a dual purpose as both a contraceptive and preventing the spread of HIV and other STDs.

MCIP: Thank you so much for your time, Dr. Sharma!

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Press coverage

The Times of India and India Edunews covered Indian work on a male contraceptive vaccine candidate which has been used with 100% success in rabbits and 85% efficacy in pygmy monkeys. If researchers have success in larger monkeys more similar to humans, toxicology studies would begin.

The local press in India and a paper for the Indian community in Thailand reported on RISUG progress based on a July 1 presentation to the Indian Medical Association. They quote RISUG's inventor Prof. Sujoy K. Guha and speculate on limited availability of this vas-based polymer injectable as early as the end of next year if the rest of the clinical trial proceeds on schedule. The journalist mixes up numbers from several different clinical trials. The interview with R.S. Sharma above contains fact-checked numbers from MCIP rather than the popular press.

India to market first male contraceptive injection in 2009
Thaindian News, 15 July
Mangalorean, 16 July

Amy Crawford's Columbia News Service story continues to be picked up by papers around the country. This well-researched piece covers the retinoic acid receptor antagonist research at Columbia University and discusses the struggles faced by hormonal researchers — in addition to mentioning suspensories, RISUG, and the IVD.

ABC News covers male contraception research (hormones and a vas clip), sacrificing accuracy for humor, and displaying a "let's make men suffer like we have" attitude. Want to have your say? Use the comments section to let people know they can find more accurate information on the subject, without the male-bashing, at MaleContraceptives.org.

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Calendar of events

Planning to attend a male contraception or andrology-related event that's not listed here? Let us know so we can post it and alert your colleagues to interesting upcoming events.

August 3-8
 Gordon Research Conferences – Reproductive Tract Biology; Andover, NH, USA
August 25-28
Endocrine Society of Australia & Society for Reproductive Biology Annual Scientific Meeting; Melbourne, VIC, Australia
September 15
Deadline for Asia-Pacific Council on Contraception abstract submission
September 26
 World Contraception Day
October 1-3
Androgens 2008, 5th Biennial Meeting on Androgen Receptor Function; Rotterdam, The Netherlands
October 15-17
 14th International Workshop on the Development and Function of the Reproductive Organs; Villa Mondragone, Italy
October 15
Deadline for International Congress of Andrology abstract submission
October 22-25
 30th Annual British Adrology Society Meeting; Gijon, Spain
October 25-28
2008 American Public Health Association Annual Meeting & Exposition; San Diego, CA, USA
November 8-12
 American Society for Reproductive Medicine 64th Annual Meeting; San Francisco, CA, USA
November 24
 Deadline for European Congress of Endocrinology abstract submission
November 26-28
5th European Congress of Andrology; Rome, Italy
December 1
 Deadline for North American Testis Workshop abstract submission
December 4-6
Second Congress of the Asia-Pacific Council on Contraception (APCOC); Macao, China
December 15
 Deadline for World Congress on Reproduction abstract submission
2009
March 5-8
 13th World Congress on Human Reproduction; Venice, Italy
March 7-10
 9th International Congress on Andrology; Barcelona, Spain
April 1-4
 20th North American Testis Workshop; Philadelphia, PA, USA
April 4-7
 34th American Society of Andrology meeting; Philadelphia, PA, USA
April 25-29
 11th European Congress of Endocinology; Istanbul, Turkey
May 31
 Deadline for Asia-Pacific Forum on Andrology abstract submission
October 10-13
 Third Asia-Pacific Forum on Andrology; Nanjing, China

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Editors

Kirsten Thompson, Director of the Male Contraception Coalition (MCC)
Email: Kirsten@MaleContraceptives.org
Phone: +1 (443) 858-1183

Elaine Lissner, Director of the Male Contraception Information Project (MCIP)
Email: Lissner@NewMaleContraception.org
Phone: +1 (415) 839-6304